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1.
mSystems ; 9(1): e0070023, 2024 Jan 23.
Article in English | MEDLINE | ID: mdl-38079111

ABSTRACT

Circadian clock arrays in multicellular filaments of the heterocyst-forming cyanobacterium Anabaena sp. strain PCC 7120 display remarkable spatio-temporal coherence under nitrogen-replete conditions. To shed light on the interplay between circadian clocks and the formation of developmental patterns, we followed the expression of a clock-controlled gene under nitrogen deprivation, at the level of individual cells. Our experiments showed that differentiation into heterocysts took place preferentially within a limited interval of the circadian clock cycle, that gene expression in different vegetative intervals along a developed filament was discoordinated, and that the circadian clock was active in individual heterocysts. Furthermore, Anabaena mutants lacking the kaiABC genes encoding the circadian clock core components produced heterocysts but failed in diazotrophy. Therefore, genes related to some aspect of nitrogen fixation, rather than early or mid-heterocyst differentiation genes, are likely affected by the absence of the clock. A bioinformatics analysis supports the notion that RpaA may play a role as master regulator of clock outputs in Anabaena, the temporal control of differentiation by the circadian clock and the involvement of the clock in proper diazotrophic growth. Together, these results suggest that under nitrogen-deficient conditions, the clock coherent unit in Anabaena is reduced from a full filament under nitrogen-rich conditions to the vegetative cell interval between heterocysts.IMPORTANCECircadian clocks, from unicellular organisms to animals, temporally align biological processes to day and night cycles. We study the dynamics of a circadian clock-controlled gene at the individual cell level in the multicellular filamentous cyanobacterium Anabaena, under nitrogen-stress conditions. Under these conditions, some cells along filaments differentiate to carry out atmospheric nitrogen fixation and lose their capability for oxygenic photosynthesis. We found that clock synchronization is limited to organismic units of contiguous photosynthetic cells, contrary to nitrogen-replete conditions in which clocks are synchronized over a whole filament. We provided evidence that the circadian clock regulates the process of differentiation, allowing it to occur preferentially within a limited time window during the circadian clock period. Lastly, we present evidence that the signal from the core clock to clock-regulated genes is conveyed in Anabaena as in unicellular cyanobacteria.


Subject(s)
Anabaena , Circadian Clocks , Cyanobacteria , Circadian Clocks/genetics , Anabaena/genetics , Cyanobacteria/metabolism , Cell Differentiation/genetics , Nitrogen/metabolism
2.
mBio ; 13(4): e0116522, 2022 08 30.
Article in English | MEDLINE | ID: mdl-35876506

ABSTRACT

Bacteria in general serve two main tasks: cell growth and division. Both processes include peptidoglycan extension to allow cell expansion and to form the poles of the daughter cells, respectively. The cyanobacterium Anabaena forms filaments of communicated cells in which the outer membrane and the peptidoglycan sacculus, which is engrossed in the intercellular regions between contiguous cells, are continuous along the filament. During the growth of Anabaena, peptidoglycan incorporation was weak at the cell periphery. During cell division, midcell peptidoglycan incorporation matched the localization of the divisome, and incorporation persisted in the intercellular septa, even after the division was completed. MreB, MreC, and MreD were located throughout the cell periphery and, in contrast to other bacteria, also to the divisome all along midcell peptidoglycan growth. In Anabaena mutants bearing inactivated mreB, mreC, or mreD genes, which showed conspicuous alterations in the filament morphology, consecutive septal bands of peptidoglycan growth were frequently not parallel to each other and were irregularly spaced along the filament, reproducing the disposition of the Z-ring. Both lateral and septal growth was impaired in strains down-expressing Z-ring components, and MreB and MreD appeared to directly interact with some divisome components. We propose that, in Anabaena, association with the divisome is a way for localization of MreB, MreC, and MreD at the cell poles, where they regulate lateral, midcell, and septal peptidoglycan growth with the latter being involved in localization and maintenance of the intercellular septal-junction protein structures that mediate cell-cell communication along the filament. IMPORTANCE Peptidoglycan surrounds the bacterial cell, being essential for the determination of the bacterium-specific morphology and survival. Peptidoglycan growth has been thoroughly investigated in some model rod-shaped bacteria, and more recently some representatives with disparate morphologies became into focus, revealing that patterns of peptidoglycan growth are much more diverse than previously anticipated. Anabaena forms filaments of communicated cells exhibiting features of multicellular organisms, such as the production of morphogens and coupled circadian oscillations. Here, we showed that Anabaena presented a distinct pattern of peptidoglycan growth characterized by continuous incorporation of material at the polar intercellular regions, contributing to assembling and maintaining the protein complexes that expand the septal peptidoglycan mediating intercellular molecular exchange in the filament.


Subject(s)
Anabaena , Peptidoglycan , Bacterial Proteins/genetics , Bacterial Proteins/metabolism , Cell Division , Cell Wall/metabolism , Peptidoglycan/metabolism
3.
Environ Microbiol ; 23(8): 4823-4837, 2021 08.
Article in English | MEDLINE | ID: mdl-34296514

ABSTRACT

FtsZ is a tubulin-like GTPase that polymerizes to initiate the process of cell division in bacteria. Heterocysts are terminally differentiated cells of filamentous cyanobacteria that have lost the capacity for cell division and in which the ftsZ gene is downregulated. However, mechanisms of FtsZ regulation during heterocyst differentiation have been scarcely investigated. The patD gene is NtcA dependent and involved in the optimization of heterocyst frequency in Anabaena sp. PCC 7120. Here, we report that the inactivation of patD caused the formation of multiple FtsZ-rings in vegetative cells, cell enlargement, and the retention of peptidoglycan synthesis activity in heterocysts, whereas its ectopic expression resulted in aberrant FtsZ polymerization and cell division. PatD interacted with FtsZ, increased FtsZ precipitation in sedimentation assays, and promoted the formation of thick straight FtsZ bundles that differ from the toroidal aggregates formed by FtsZ alone. These results suggest that in the differentiating heterocysts, PatD interferes with the assembly of FtsZ. We propose that in Anabaena FtsZ is a bifunctional protein involved in both vegetative cell division and regulation of heterocyst differentiation. In the differentiating cells PatD-FtsZ interactions appear to set an FtsZ activity that is insufficient for cell division but optimal to foster differentiation.


Subject(s)
Anabaena , Cyanobacteria , Anabaena/genetics , Anabaena/metabolism , Bacterial Proteins/genetics , Bacterial Proteins/metabolism , Cell Division/genetics , Cyanobacteria/metabolism , Gene Expression Regulation, Bacterial
4.
Elife ; 102021 03 22.
Article in English | MEDLINE | ID: mdl-33749592

ABSTRACT

Circadian clocks display remarkable reliability despite significant stochasticity in biomolecular reactions. We study the dynamics of a circadian clock-controlled gene at the individual cell level in Anabaena sp. PCC 7120, a multicellular filamentous cyanobacterium. We found significant synchronization and spatial coherence along filaments, clock coupling due to cell-cell communication, and gating of the cell cycle. Furthermore, we observed low-amplitude circadian oscillatory transcription of kai genes encoding the post-transcriptional core oscillatory circuit and high-amplitude oscillations of rpaA coding for the master regulator transducing the core clock output. Transcriptional oscillations of rpaA suggest an additional level of regulation. A stochastic one-dimensional toy model of coupled clock cores and their phosphorylation states shows that demographic noise can seed stochastic oscillations outside the region where deterministic limit cycles with circadian periods occur. The model reproduces the observed spatio-temporal coherence along filaments and provides a robust description of coupled circadian clocks in a multicellular organism.


Subject(s)
Anabaena/genetics , Cell Communication , Circadian Clocks/genetics , Anabaena/cytology , Anabaena/metabolism , Cell Cycle
5.
Braz. dent. sci ; 24(4, suppl 1): 1-7, 2021. tab, ilus
Article in English | LILACS, BBO - Dentistry | ID: biblio-1349350

ABSTRACT

Objective: Despite the increase in the female contribution to careers in the health sector, Dentistry has shown slow progress towards gender equality. The objective of this study was to quantify the proportion of women in the editorial committees of dental journals in the world. Material and Methods: Dental journals published in the world were compiled, which met inclusion criteria: dental journals indexed to Scopus in their 2020 edition, access to the composition of the editorial committee. Non-current journals, without access to their website, journals not classified in a quartile, and journals with publishers outside their country of origin, were excluded. The selection of journals was carried out from January 11 to 19, 2021. The analysis variables were the composition of the editorial committee, dental specialty according to the SJR category and the title of the journal, quartile of the journal, and country of origin of the editorial headquarters. Results: One hundred eighty nine journals were identified. Women represented 22.91% for the position of director or editor-in-chief. With respect to associate editors and members of the editorial board, 24.76% and 22.91% were women, respectively. Likewise, greater female participation was observed in Q2 and Q1 journals and in thematic areas of Geriatric Dentistry, Dental Education, Dental Public Health, and Basic Sciences. Conclusion: The findings demonstrate the low proportion of women on the editorial boards of dental journals in the world (AU)


Objetivo: Apesar do aumento da contribuição feminina para as carreiras no setor da saúde, a odontologia tem apresentado avanços lentos em direção à equidade de gênero. O objetivo deste estudo foi quantificar a proporção de mulheres nos comitês editoriais de periódicos odontológicos no mundo. Material e Métodos: Foram compilados periódicos odontológicos publicados no mundo, que atenderam aos critérios de inclusão: periódicos odontológicos indexados ao Scopus em sua edição de 2020, acesso à composição do comitê editorial. Foram excluídos os periódicos não atuais, sem acesso ao site, os periódicos não classificados em quartil e os periódicos com editoras fora do país de origem. A seleção dos periódicos foi realizada no período de 11 a 19 de janeiro de 2021. As variáveis de análise foram a composição do comitê editorial, especialidade odontológica de acordo com a categoria SJR e o título do periódico, quartil do periódico e país de origem da sede do editorial. Resultados:Cento e oitenta e nove periódicos foram identificados. As mulheres representaram 22,91% para o cargo de diretora ou redatora-chefe. Em relação aos editores associados e membros do comitê editorial, 24,76% e 22,91% eram mulheres, respectivamente. Da mesma forma, foi observada maior participação feminina nos periódicos Q2 e Q1 e nas áreas temáticas de Odontologia Geriátrica, Educação Odontológica, Saúde Pública Odontológica e Ciências Básicas. Conclusão: Os achados demonstram a baixa proporção de mulheres nos conselhos editoriais de periódicos odontológicos no mundo. (AU)


Subject(s)
Women , Dentistry , Gender Identity
6.
mSphere ; 5(3)2020 05 20.
Article in English | MEDLINE | ID: mdl-32434840

ABSTRACT

The Anabaena organismic unit is a filament of communicating cells. Under conditions of nitrogen scarcity, some cells along the filament differentiate into heterocysts, which are specialized in the fixation of atmospheric N2 and provide the vegetative cells with N2 fixation products. At a certain stage, the differentiation process becomes irreversible, so that even when nitrogen is replenished, no return to the vegetative cell state takes place, possibly as a consequence of loss of cell division capacity. Upon N-stepdown, midcell FtsZ-rings were detected in vegetative cells, but not in differentiating cells, and this was also the case for ZipN, an essential protein that participates in FtsZ tethering to the cytoplasmic membrane and divisome organization. Later, expression of ftsZ was arrested in mature heterocysts. PatA is a protein required for the differentiation of intercalary heterocysts in Anabaena The expression level of the patA gene was increased in differentiating cells, and a mutant strain lacking PatA exhibited enhanced FtsZ-rings. PatA was capable of direct interactions with ZipN and SepF, another essential component of the Anabaena Z-ring. Thus, PatA appears to promote inhibition of cell division in the differentiating cells, allowing progress of the differentiation process. PatA, which in mature heterocysts was detected at the cell poles, could interact also with SepJ, a protein involved in production of the septal junctions that provide cell-cell adhesion and intercellular communication in the filament, hinting at a further role of PatA in the formation or stability of the intercellular structures that are at the basis of the multicellular character of AnabaenaIMPORTANCEAnabaena is a cyanobacterial model that represents an ancient and simple form of biological multicellularity. The Anabaena organism is a filament of cohesive and communicating cells that can include cells specialized in different tasks. Thus, under conditions of nitrogen scarcity, certain cells of the filament differentiate into heterocysts, which fix atmospheric nitrogen and provide organic nitrogen to the rest of cells, which, in turn, provide heterocysts with organic carbon. Heterocyst differentiation involves extensive morphological, biochemical, and genetic changes, becoming irreversible at a certain stage. We studied the regulation during heterocyst differentiation of several essential components of the Anabaena cell division machinery and found that protein PatA, which is required for differentiation and is induced in differentiating cells, interacts with essential cell division factors and destabilizes the cell division complex. This suggests a mechanism for establishment of commitment to differentiation by inhibition of cell division.


Subject(s)
Anabaena/genetics , Bacterial Proteins/genetics , Cell Division/genetics , Gene Expression Regulation, Bacterial , Anabaena/physiology , Bacterial Proteins/metabolism
7.
Sci Rep ; 9(1): 2744, 2019 02 26.
Article in English | MEDLINE | ID: mdl-30808920

ABSTRACT

The organismic unit of heterocyst-forming cyanobacteria is a filament of communicating cells connected by septal junctions, proteinaceous structures bridging the cytoplasms of contiguous cells. This distinct bacterial organization is preserved during cell division. In Anabaena, deletion of the zipN gene could not be segregated. We generated strain CSL109 that expresses zipN from a synthetic regulatable promoter. Under conditions of ZipN depletion, cells progressively enlarged, reflecting restricted cell division, and showed drastic morphological alterations including cell detachment from the filaments, to finish lysing. In contrast to the wild-type localization in midcell Z-rings, FtsZ was found in delocalized aggregates in strain CSL109. Consistently, the proportion of membrane-associated to soluble FtsZ in fractionated cell extracts was lower in CSL109. Bacterial two-hybrid analysis showed that ZipN interacts with FtsZ and other cell-division proteins including cytoplasmic Ftn6 and SepF, and polytopic FtsW, FtsX, FtsQ and FtsI. Additionally, ZipN interacted with the septal protein SepJ, and in CSL109 depletion of ZipN was concomitant with a progressive loss of septal specificity of SepJ. Thus, in Anabaena ZipN represents an essential FtsZ membrane tether and an organizer of the divisome, and it contributes to the conformation of septal structures for filament integrity and intercellular communication.


Subject(s)
Bacterial Proteins/metabolism , Cell Membrane/metabolism , Cyanobacteria/growth & development , Cyanobacteria/metabolism , Cytoplasm/metabolism , Gene Expression Regulation, Bacterial , Bacterial Proteins/genetics
8.
Mol Microbiol ; 111(4): 883-897, 2019 04.
Article in English | MEDLINE | ID: mdl-30636068

ABSTRACT

Arginine participates widely in metabolic processes. The heterocyst-forming cyanobacterium Anabaena catabolizes arginine to produce proline and glutamate, with concomitant release of ammonium, as major products. Analysis of mutant Anabaena strains showed that this catabolic pathway is the product of two genes, agrE (alr4995) and putA (alr0540). The predicted PutA protein is a conventional, bifunctional proline oxidase that produces glutamate from proline. In contrast, AgrE is a hitherto unrecognized enzyme that contains both an N-terminal α/ß propeller domain and a unique C-terminal domain of previously unidentified function. In vitro analysis of the proteins expressed in Escherichia coli or Anabaena showed arginine dihydrolase activity of the N-terminal domain and ornithine cyclodeaminase activity of the C-terminal domain, overall producing proline from arginine. In the diazotrophic filaments of Anabaena, ß-aspartyl-arginine dipeptide is transferred from the heterocysts to the vegetative cells, where it is cleaved producing aspartate and arginine. Both agrE and putA were found to be expressed at higher levels in vegetative cells than in heterocysts, implying that arginine is catabolized by the AgrE-PutA pathway mainly in the vegetative cells. Expression in Anabaena of a homolog of the C-terminal domain of AgrE obtained from Methanococcus maripaludis enabled us to identify an archaeal ornithine cyclodeaminase.


Subject(s)
Ammonia-Lyases/metabolism , Anabaena/enzymology , Arginine/metabolism , Proline/metabolism , Ammonia-Lyases/genetics , Anabaena/genetics , Bacterial Proteins/genetics , Bacterial Proteins/metabolism , Escherichia coli/genetics , Escherichia coli/metabolism , Gene Expression Regulation, Bacterial , Membrane Proteins/genetics , Membrane Proteins/metabolism , Metabolic Networks and Pathways , Nitrogen Fixation , Proline Oxidase/genetics , Proline Oxidase/metabolism
9.
Biochim Biophys Acta Gene Regul Mech ; 1862(7): 673-684, 2019 07.
Article in English | MEDLINE | ID: mdl-29719238

ABSTRACT

Filamentous, heterocyst-forming cyanobacteria are among the simplest multicellular systems in Nature. In the absence of combined nitrogen, the filaments consist of vegetative cells that fix CO2 through oxygenic photosynthesis and micro-oxic heterocysts specialized for the fixation of N2 in a proportion of about 10 to 1. The development of a heterocyst-containing filament involves differentiation of vegetative cells into heterocysts in a process that requires a distinct gene expression program. Two transcription factors are strictly required, NtcA and HetR. The CRP-family protein NtcA directly activates the expression of multiple genes during heterocyst differentiation - in some cases assisted by coactivators including HetR - and in mature heterocysts, whereas HetR is needed to build high NtcA levels in differentiating heterocysts and directly activates some particular genes. A few other regulators of gene expression participate at specific differentiation steps, and a specific transcription factor, CnfR, activates nif gene expression under the micro-oxic conditions of the heterocyst.


Subject(s)
Bacterial Proteins/genetics , Cyanobacteria/growth & development , Transcription Factors/genetics , Bacterial Proteins/chemistry , Cyanobacteria/genetics , Cyanobacteria/metabolism , Gene Expression Regulation, Bacterial , Gene Expression Regulation, Developmental , Models, Molecular , Transcription Factors/chemistry
10.
Front Microbiol ; 9: 2260, 2018.
Article in English | MEDLINE | ID: mdl-30333801

ABSTRACT

Filamentous cyanobacteria grow by intercalary cell division, which should involve distinct steps compared to those producing separate daughter cells. The N-terminal region of FtsZ is highly conserved in the clade of filamentous cyanobacteria capable of cell differentiation. A derivative of the model strain Anabaena sp. PCC 7120 expressing only an FtsZ lacking the amino acids 2-51 of the N-terminal peptide (ΔN-FtsZ) could not be segregated. Strain CSL110 expresses both ΔN-FtsZ, from the endogenous ftsZ gene promoter, and the native FtsZ from a synthetic regulated promoter. Under conditions of ΔN-FtsZ predominance, cells of strain CSL110 progressively enlarge, reflecting reduced cell division, and show instances of asymmetric cell division and aberrant Z-structures notably differing from the Z-ring formed by FtsZ in the wild type. In bacterial 2-hybrid assays FtsZ interacted with ΔN-FtsZ. However, ΔN-FtsZ-GFP appeared impaired for incorporation into Z-rings when expressed together with FtsZ. FtsZ, but not ΔN-FtsZ, interacted with the essential protein SepF. Both FtsZ and ΔN-FtsZ polymerize in vitro exhibiting comparable GTPase activities. However, filaments of FtsZ show a distinct curling forming toroids, whereas ΔN-FtsZ form thick bundles of straight filaments. Thus, the N-terminal FtsZ sequence appears to contribute to a distinct FtsZ polymerization mode that is essential for cell division and division plane location in Anabaena.

11.
Plant Cell Physiol ; 58(2): 256-265, 2017 02 01.
Article in English | MEDLINE | ID: mdl-28007969

ABSTRACT

All known cyanobacteria contain Cyt c6, a small soluble electron carrier protein whose main function is to transfer electrons from the Cyt b6f complex to PSI, although it is also involved in respiration. We have previously described a second isoform of this protein, the Cyt c6-like, whose function remains unknown. Here we describe a third isoform of Cyt c6 (here called Cytc6-3), which is only found in heterocyst-forming filamentous cyanobacteria. Cyt c6-3 is expressed in vegetative cells but is specifically repressed in heterocysts cells under diazotrophic growth conditions. Although there is a close structural similarity between Cyt c6-3 and Cyt c6 related to the general protein folding, Cyt c6-3 presents differential electrostatic surface features as compared with Cyt c6, its expression is not copper dependent and has a low reactivity towards PSI. According to the different expression pattern, functional reactivity and structural properties, Cyt c6-3 has to play an as yet to be defined regulatory role related to heterocyst differentiation.


Subject(s)
Bacterial Proteins/metabolism , Cyanobacteria/metabolism , Protein Isoforms/metabolism , Electron Transport/physiology , Photosynthesis/physiology , Plastocyanin/metabolism
12.
Mol Microbiol ; 99(4): 808-19, 2016 Feb.
Article in English | MEDLINE | ID: mdl-26552991

ABSTRACT

Many filamentous cyanobacteria respond to the external cue of nitrogen scarcity by the differentiation of heterocysts, cells specialized in the fixation of atmospheric nitrogen in oxic environments. Heterocysts follow a spatial pattern along the filament of two heterocysts separated by ca. 10-15 vegetative cells performing oxygenic photosynthesis. HetR is a transcriptional regulator that directs heterocyst differentiation. In the model strain Anabaena sp. PCC 7120, the HetR protein was observed in various oligomeric forms in vivo, including a tetramer that peaked with maximal hetR expression during differentiation. Tetramers were not detected in a hetR point mutant incapable of differentiation, but were conspicuous in an over-differentiating strain lacking the PatS inhibitor. In differentiated filaments the HetR tetramer was restricted to heterocysts, being undetectable in vegetative cells. HetR co-purified with RNA polymerase from Anabaena mainly as a tetramer. In vitro, purified recombinant HetR was distributed between monomers, dimers, trimers and tetramers, and it was phosphorylated when incubated with (γ-(32)P)ATP. Phosphorylation and PatS hampered the accumulation of HetR tetramers and impaired HetR binding to DNA. In summary, tetrameric HetR appears to represent a functionally relevant form of HetR, whose abundance in the Anabaena filament could be negatively regulated by phosphorylation and by PatS.


Subject(s)
Anabaena/genetics , Bacterial Proteins/chemistry , Bacterial Proteins/metabolism , Gene Expression Regulation, Bacterial , Transcription Factors/chemistry , Transcription Factors/metabolism , Anabaena/metabolism , Bacterial Proteins/genetics , Nitrogen/metabolism , Phosphorylation , Point Mutation , Recombinant Proteins/chemistry , Recombinant Proteins/isolation & purification , Recombinant Proteins/metabolism , Transcription Factors/genetics
13.
Rev Med Inst Mex Seguro Soc ; 53 Suppl 2: S212-7, 2015.
Article in Spanish | MEDLINE | ID: mdl-26462519

ABSTRACT

Despite the use of the screening test, such as Papanicolaou, and the detection of human papillomavirus (HPV), cervical cancer remains as a public health problem in México and it is the second leading cause of death for malignant neoplasias among women. High-risk HPV infection is the main risk factor for the development of premalignant lesions and cervical cancer; however, HPV infection is not the only factor; there are various genetic and epigenetic alterations required for the development of neoplasias; some of them have been described and even in some cases they have been suggested as biomarkers for prognosis. However, in contrast with other cancer types, such as breast cancer, in cervical cancer the use of biomarkers has not been established for clinical applications. Unlike genetic alterations, epigenetic alterations are potentially reversible; in this sense, their characterization is important, since they have not only a potential use as biomarkers, but they also could represent new therapeutic targets for treatment of cervical cancer. This review describes some of the more common epigenetic alterations in cervical cancer and its potential use in routine clinical practice.


A pesar del empleo de los métodos de tamizaje como el Papanicolaou y la detección de virus del papiloma humano (VPH), el cáncer cervicouterino (CaCU) sigue siendo un problema de salud pública en México, ya que es la segunda causa de muerte por neoplasias malignas en mujeres. La infección por VPH de alto potencial oncogénico es el principal factor de riesgo para el desarrollo de lesiones precursoras y CaCU; sin embargo, no es suficiente, dado que se requiere de diversas alteraciones genéticas y epigenéticas para el desarrollo de la neoplasia y un gran número de ellas han sido descritas, incluso en algunos casos se ha sugerido su uso como biomarcadores en la progresión o predictivos de respuesta. A diferencia de lo que sucede con otros tipos de cáncer, como el de mama, en el CaCU no se ha establecido el uso de marcadores para su aplicación en la clínica. A diferencia de las alteraciones genéticas, las alteraciones epigenéticas son potencialmente reversibles y su caracterización no solo tiene potencial para el uso como biomarcadores, sino como blancos de nuevas terapias. En esta revisión se describen algunas de las alteraciones epigenéticas más características en el CaCU, con potencial uso para la clínica.


Subject(s)
Epigenesis, Genetic , Uterine Cervical Neoplasms/genetics , DNA Methylation , Disease Progression , Female , Genetic Markers , Genetic Testing , Histone Code , Humans , RNA, Untranslated , Uterine Cervical Neoplasms/diagnosis , Uterine Cervical Neoplasms/therapy
14.
FEBS Lett ; 588(9): 1787-94, 2014 May 02.
Article in English | MEDLINE | ID: mdl-24685693

ABSTRACT

In Anabaena, the pipX gene is induced in the cells differentiating into heterocysts, being the PipX factor required for full expression of late heterocyst-specific genes. Here we show that PipX has a positive effect on in vitro binding of the transcription factor NtcA to DNA, as well as on transcript production, in different NtcA-dependent promoters. We found that the cox3 operon, encoding a heterocyst-specific terminal respiratory oxidase, is expressed from three nitrogen-regulated promoters to which NtcA binds. At the three sites, NtcA binding is potentiated by PipX. Thus, PipX has a direct effect on gene expression influencing the activity of NtcA.


Subject(s)
Anabaena/genetics , Bacterial Proteins/genetics , Electron Transport Complex IV/genetics , Transcription Factors/genetics , Bacterial Proteins/chemistry , Base Sequence , Consensus Sequence , DNA Footprinting , DNA, Bacterial/chemistry , DNA, Bacterial/genetics , Gene Expression Regulation, Bacterial , Genes, Bacterial , Molecular Sequence Data , Promoter Regions, Genetic , Protein Binding , Transcription Factors/chemistry , Transcription, Genetic
15.
FEBS Lett ; 587(16): 2682-90, 2013 Aug 19.
Article in English | MEDLINE | ID: mdl-23851073

ABSTRACT

In Anabaena sp. PCC 7120, FurA is a global transcriptional regulator whose expression is strongly induced by NtcA in proheterocysts and remains stably expressed in mature heterocysts. In the present study, overexpression of furA partially suppressed heterocyst differentiation by impairing morphogenesis at an early stage. Recombinant purified FurA specifically bound in vitro to the promoter regions of ntcA, while quantitative RT-PCR analyses indicated that furA overexpression strongly affected the transient increase of ntcA expression that occurs shortly after nitrogen step-down. Overall, the results suggest a connection between iron homeostasis and heterocyst differentiation via FurA, by modulating the expression of ntcA.


Subject(s)
Anabaena/cytology , Bacterial Proteins/genetics , Gene Expression Regulation, Bacterial , Iron/metabolism , Base Sequence , Cell Differentiation , Deoxyribonuclease I/metabolism , Deoxyribonucleases/chemistry , Homeostasis , Molecular Sequence Data , Promoter Regions, Genetic
16.
Environ Microbiol ; 14(12): 3175-87, 2012 Dec.
Article in English | MEDLINE | ID: mdl-23066898

ABSTRACT

Knowledge on the regulatory mechanisms controlling iron homeostasis in cyanobacteria is limited. In Anabaena sp. PCC 7120, the ferric uptake regulator FurA is a constitutive and essential protein whose expression is induced under iron deprivation. Our previous analyses have shown that this protein acts as a global transcriptional regulator, controlling the expression of several genes belonging to different functional categories, including schT, a gene coding for a TonB-dependent schizokinen transporter. In the present study we analysed the impact of FurA overexpression and iron availability on the transcriptional modulation of a broad range of Anabaena iron uptake, transport, storage and cellular iron utilization mechanisms, including enzymes involved in siderophore biosynthesis, TonB-dependent siderophore outer membrane transporters, siderophore periplasmic binding proteins, ABC inner membrane permeases, ferritin Dps family proteins, and enzymes involved in tetrapyrrole biosynthesis. By combining reverse transcription-PCR analyses, electrophoretic mobility shift assays and DNase I footprinting experiments, we defined a variety of novel direct iron-dependent transcriptional targets of this metalloregulator, including genes encoding at least five enzymes involved in the tetrapyrrole biosynthesis pathway. The results unravel the role of FurA as the master regulator of iron homeostasis in Anabaena sp. PCC 7120, providing new insights into the Fur regulons in cyanobacteria.


Subject(s)
Anabaena/genetics , Anabaena/metabolism , Bacterial Proteins/metabolism , Homeostasis/genetics , Iron/metabolism , Regulon/physiology , Tetrapyrroles/biosynthesis , Binding Sites , Biological Transport/genetics , Heme/biosynthesis , Heme/metabolism , Hydroxamic Acids/metabolism , Membrane Transport Proteins/metabolism , Promoter Regions, Genetic , Siderophores/biosynthesis , Siderophores/metabolism
17.
Adicciones ; 24(2): 161-72, 2012.
Article in Spanish | MEDLINE | ID: mdl-22648319

ABSTRACT

Several neuropsychological studies have shown that chronic cannabis users have cognitive impairments, including decision-making process. Therefore, this study aims to evaluate the process, through the somatic marker hypothesis in a sample of 41 cannabis users compared with a control group of equal size, and to analyze the influence of age, sex, education level, age of onset and amount of daily consumption. In order to do that, the software "Cartas" (similar to the Iowa Gambling Task), was used, implementing its two versions: normal and reverse. The results show significant differences between cannabis users and control group in the normal and reverse task execution. By block analysis, the control group obtained higher scores in the normal task execution, however, in the reverse task, the differences between groups are present in the initial task execution but not final task execution. None of the analyzed variables (age, sex ...) are significantly related to task performance. These results suggest the existence of alterations in the decision making process of consumers cannabis, which may relate to the difficulty in generating somatic markers, and not for insensitivity punishments insensitivity.


Subject(s)
Decision Making , Marijuana Abuse/psychology , Adult , Female , Humans , Male , Middle Aged , Young Adult
18.
Adicciones (Palma de Mallorca) ; 24(2): 161-172, abr.-jun. 2012. ilus, tab, graf
Article in Spanish | IBECS | ID: ibc-101465

ABSTRACT

Diversos estudios neuropsicológicos han demostrado que los consumidores crónicos de cannabis presentan deterioros cognitivos, incluyendo el proceso de toma de decisiones. Por ello, el presente estudio tiene como objetivo evaluar dicho proceso, a través de la hipótesis del marcador somático, en una muestra de 41 consumidores de cannabis comparándolo con un grupo control de igual tamaño, así como analizarla influencia de la edad, sexo, nivel de estudios, edad de inicio y cantidad de consumo diario. Para ello, se ha utilizado el programa "Cartas" (similar a la Iowa Gambling Task), administrando dos versiones: normal e inversa. Los resultados muestran diferencias estadísticamente significativas entre los consumidores de cannabis y el grupo control en la ejecución de la tarea normal e inversa. En el análisis por bloques, el grupo control obtiene puntuaciones superiores en la ejecución de la tarea normal, sin embargo, en la tarea inversa, las diferencias entre ambos grupos se dan en la parte inicial pero no en la final. Ninguna de las variables analizadas (edad, sexo,...) se relaciona significativamente con el rendimiento de la tarea. Estos resultados sugieren la existencia de alteraciones en el proceso de toma de decisiones de los consumidores de cannabis, que pueden relacionarse con la dificultad para generar marcadores somáticos, y no con la insensibilidad a las pérdidas(AU)


Several neuropsychological studies have shown that chronic cannabis users have cognitive impairments, including decision-making process. Therefore, this study aims to evaluate the process, through the somatic marker hypothesis in a sample of 41 cannabis users compared with a control group of equal size, and to analyze the influence of age, sex, education level, age of onset and amount of daily consumption. In order to do that, the software "Cartas" (similar to the Iowa Gambling Task),was used, implementing its two versions: normal and reverse. The results show significant differences between cannabis users and control group in the normal and reverse task execution. By block analysis, the control group obtained higher scores in the normal task execution, however, in the reverse task, the differences between groups are present in the initial task execution but not final task execution. None of the analyzed variables (age, sex ...) are significantly related to task performance. These results suggest the existence of alterations in the decision making process of consumers cannabis, which may relate to the difficulty in generating somatic markers, and not for insensitivity punishments insensitivity(AU)


Subject(s)
Humans , Male , Female , Adult , Middle Aged , Marijuana Smoking/legislation & jurisprudence , Decision Making/ethics , Marijuana Smoking/prevention & control , Marijuana Smoking/psychology , Marijuana Smoking/trends , Marijuana Smoking/therapy , Decision Making , Decision Making/physiology
19.
J Bacteriol ; 194(11): 2939-48, 2012 Jun.
Article in English | MEDLINE | ID: mdl-22467790

ABSTRACT

Heterocyst differentiation is orchestrated by the N control transcriptional regulator NtcA and the differentiation-specific factor HetR. In Anabaena sp. strain PCC 7120, the devBCA operon is expressed from two different promoters activated upon N stepdown. The distal devB promoter (transcription start point [TSP] located at position -704) represents a canonical class II NtcA-activated promoter, including a consensus NtcA-binding site centered 39.5 nucleotides upstream from the TSP. Transcription activation from a second TSP (-454) requires NtcA and is impaired in hetR mutants. In a wild-type background, three different DNA fragments, including both or each individual promoter, directed gfp expression localized mainly to proheterocysts and heterocysts. Expression was undetectable in an ntcA background and, for the fragment including the proximal promoter alone, also in a hetR background. In spite of the absence of consensus NtcA-binding sequences between the two TSPs, NtcA was shown to interact with this DNA region, and NtcA and its effector, 2-oxoglutarate, were necessary and sufficient for in vitro transcription from the -454 TSP. No HetR binding to the DNA or in vitro transcription from the proximal devB TSP promoted by HetR alone were detected. However, a moderate positive effect of HetR on NtcA binding to the DNA between the two devB TSPs was observed. The proximal devB promoter appears to represent a suboptimal NtcA-activated promoter for which HetR may act as a coactivator, with the physiological effect of restricting gene activation to conditions of prevalence of high NtcA and HetR levels, such as those taking place during heterocyst differentiation.


Subject(s)
Anabaena/growth & development , Bacterial Proteins/metabolism , Gene Expression Regulation, Bacterial , Promoter Regions, Genetic , Trans-Activators/metabolism , Transcriptional Activation , Anabaena/genetics , Anabaena/metabolism , Bacterial Proteins/chemistry , Bacterial Proteins/genetics , Binding Sites , Gene Expression Regulation, Developmental , Operon , Protein Binding , Trans-Activators/chemistry , Trans-Activators/genetics
20.
Rev Gaucha Enferm ; 32(3): 443-50, 2011 Sep.
Article in Portuguese | MEDLINE | ID: mdl-22165388

ABSTRACT

This study aimed to evaluate and compare the development of hospitalized children before and after art therapy interventions. Qualitative case studies were undertaken in this descriptive-exploratory research, based on the developmental evaluation of the children. The study participants were five children between seven and ten years old, in the Hospital of Tropical Illnesses (HDT) in the city of Goiânia, state of Goiás, Brazil, in 2006. Results showed that art therapy interventions efficiently promoted children's development. Art therapy is a resource for positively channeling the variables of hospitalized children's development and for neutralizing affective factors that naturally appear, as well as for exposing the child's healthier potentials, which sometimes receive little stimulus in the context of hospitalization.


Subject(s)
Art Therapy , Child Development , Child, Hospitalized/psychology , Hospitalization , Child , Female , Humans , Male
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