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1.
J Cell Sci ; 136(22)2023 11 15.
Artigo em Inglês | MEDLINE | ID: mdl-37902010

RESUMO

The contractile vacuole complex (CVC) is a dynamic and morphologically complex membrane organelle, comprising a large vesicle (bladder) linked with a tubular reticulum (spongiome). CVCs provide key osmoregulatory roles across diverse eukaryotic lineages, but probing the mechanisms underlying their structure and function is hampered by the limited tools available for in vivo analysis. In the experimentally tractable ciliate Tetrahymena thermophila, we describe four proteins that, as endogenously tagged constructs, localize specifically to distinct CVC zones. The DOPEY homolog Dop1p and the CORVET subunit Vps8Dp localize both to the bladder and spongiome but with different local distributions that are sensitive to osmotic perturbation, whereas the lipid scramblase Scr7p colocalizes with Vps8Dp. The H+-ATPase subunit Vma4 is spongiome specific. The live imaging permitted by these probes revealed dynamics at multiple scales including rapid exchange of CVC-localized and soluble protein pools versus lateral diffusion in the spongiome, spongiome extension and branching, and CVC formation during mitosis. Although the association with DOP1 and VPS8D implicate the CVC in endosomal trafficking, both the bladder and spongiome might be isolated from bulk endocytic input.


Assuntos
Tetrahymena thermophila , Vacúolos , Vacúolos/metabolismo , Endossomos , Proteínas/metabolismo , Mitose
2.
PLoS Biol ; 18(8): e3000756, 2020 08.
Artigo em Inglês | MEDLINE | ID: mdl-32745139

RESUMO

Recognition of self and nonself is important for outcrossing organisms, and different mating types establish the barrier against self-mating. In the unicellular ciliate T. thermophila, mating type determination requires complex DNA rearrangements at a single mat locus during conjugation to produce a type-specific gene pair (MTA and MTB) for 1 of 7 possible mating types. Surprisingly, we found that decreased expression of the DNA breakage-repair protein Ku80 at late stages of conjugation generated persistent selfing phenotype in the progeny. DNA analysis revealed multiple mating-type gene pairs as well as a variety of mis-paired, unusually arranged mating-type genes in these selfers that resemble some proposed rearrangement intermediates. They are found also in normal cells during conjugation and are lost after 10 fissions but are retained in Ku mutants. Silencing of TKU80 or TKU70-2 immediately after conjugation also generated selfing phenotype, revealing a hidden DNA rearrangement process beyond conjugation. Mating reactions between the mutant and normal cells suggest a 2-component system for self-nonself-recognition through MTA and MTB genes.


Assuntos
DNA de Protozoário/genética , Rearranjo Gênico , Autoantígeno Ku/genética , Proteínas de Protozoários/genética , Tetrahymena thermophila/genética , Conjugação Genética , Cruzamentos Genéticos , DNA de Protozoário/metabolismo , Expressão Gênica , Inativação Gênica , Autoantígeno Ku/metabolismo , Fenótipo , Proteínas de Protozoários/metabolismo , Reprodução , Tetrahymena thermophila/metabolismo
3.
Sci Rep ; 9(1): 13669, 2019 09 20.
Artigo em Inglês | MEDLINE | ID: mdl-31541152

RESUMO

Ciliates are unicellular eukaryotes known for their cellular complexity and wide range of natural habitats. How they adapt to their niches and what roles they play in ecology remain largely unknown. The genus Tetrahymena is among the best-studied groups of ciliates and one particular species, Tetrahymena thermophila, is a well-known laboratory model organism in cell and molecular biology, making it an excellent candidate for study in protist ecology. Here, based on cytochrome c oxidase subunit I (COX1) gene barcoding, we identify a total of 19 different putative Tetrahymena species and two closely related Glaucoma lineages isolated from distinct natural habitats, of which 13 are new species. These latter include 11 Tetrahymena species found in the bladder traps of Utricularia plants, the most species-rich and widely distributed aquatic carnivorous plant, thus revealing a previously unknown but significant symbiosis of Tetrahymena species living among the microbial community of Utricularia bladder traps. Additional species were collected using an artificial trap method we have developed. We show that diverse Tetrahymena species may live even within the same habitat and that their populations are highly dynamic, suggesting that the diversity and biomass of species worldwide is far greater than currently appreciated.


Assuntos
Ciclo-Oxigenase 1/genética , Lamiales/parasitologia , Análise de Sequência de DNA/métodos , Tetrahymena/classificação , Organismos Aquáticos/parasitologia , Carnivoridade , Código de Barras de DNA Taxonômico , Filogenia , Filogeografia , Proteínas de Protozoários/genética , Simbiose , Taiwan , Tetrahymena/genética , Tetrahymena/isolamento & purificação
5.
Nucleic Acids Res ; 47(10): 5181-5192, 2019 06 04.
Artigo em Inglês | MEDLINE | ID: mdl-30918956

RESUMO

Eukaryotic cells pack their genomic DNA into euchromatin and heterochromatin. Boundaries between these domains have been shown to be set by boundary elements. In Tetrahymena, heterochromatin domains are targeted for deletion from the somatic nuclei through a sophisticated programmed DNA rearrangement mechanism, resulting in the elimination of 34% of the germline genome in ∼10,000 dispersed segments. Here we showed that most of these deletions occur consistently with very limited variations in their boundaries among inbred lines. We identified several potential flanking regulatory sequences, each associated with a subset of deletions, using a genome-wide motif finding approach. These flanking sequences are inverted repeats with the copies located at nearly identical distances from the opposite ends of the deleted regions, suggesting potential roles in boundary determination. By removing and testing two such inverted repeats in vivo, we found that the ability for boundary maintenance of the associated deletion were lost. Furthermore, we analyzed the deletion boundaries in mutants of a known boundary-determining protein, Lia3p and found that the subset of deletions that are affected by LIA3 knockout contained common features of flanking regulatory sequences. This study suggests a common mechanism for setting deletion boundaries by flanking inverted repeats in Tetrahymena thermophila.


Assuntos
DNA de Protozoário/genética , Deleção de Genes , Heterocromatina/química , Proteínas de Protozoários/genética , Tetrahymena thermophila/genética , Motivos de Aminoácidos , Núcleo Celular/metabolismo , DNA de Protozoário/metabolismo , Eucromatina/química , Regulação da Expressão Gênica , Rearranjo Gênico , Genoma de Protozoário , Domínios Proteicos
6.
Front Microbiol ; 8: 189, 2017.
Artigo em Inglês | MEDLINE | ID: mdl-28232825

RESUMO

Although the presence of endosymbiotic rickettsial bacteria, specifically Candidatus Megaira, has been reported in diverse habitats and a wide range of eukaryotic hosts, it remains unclear how broadly Ca. Megaira are distributed in a single host species. In this study we seek to address whether Ca. Megaira are present in most, if not all isolates, of the parasitic ciliate Ichthyophthirius multifiliis. Conserved regions of bacterial 16S rRNA genes were either PCR amplified, or assembled from deep sequencing data, from 18 isolates/populations of I. multifiliis sampled worldwide (Brazil, Taiwan, and USA). We found that rickettsial rRNA sequences belonging to three out of four Ca. Megaira subclades could be consistently detected in all I. multifiliis samples. I. multifiliis collected from local fish farms tend to be inhabited by the same subclade of Ca. Megaira, whereas those derived from pet fish are often inhabited by more than one subclade of Ca. Megaira. Distributions of Ca. Megaira in I. multifiliis thus better reflect the travel history, but not the phylogeny, of I. multifiliis. In summary, our results suggest that I. multifiliis may be dependent on this endosymbiotic relationship, and the association between Ca. Megaira and I. multifiliis is more diverse than previously thought.

7.
PLoS Genet ; 12(11): e1006403, 2016 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-27806059

RESUMO

The maintenance of chromosome integrity is crucial for genetic stability. However, programmed chromosome fragmentations are known to occur in many organisms, and in the ciliate Tetrahymena the five germline chromosomes are fragmented into hundreds of minichromosomes during somatic nuclear differentiation. Here, we showed that there are different fates of these minichromosomes after chromosome breakage. Among the 326 somatic minichromosomes identified using genomic data, 50 are selectively eliminated from the mature somatic genome. Interestingly, many and probably most of these minichromosomes are eliminated during the growth period between 6 and 20 doublings right after conjugation. Genes with potential conjugation-specific functions are found in these minichromosomes. This study revealed a new mode of programmed DNA elimination in ciliates similar to those observed in parasitic nematodes, which could play a role in developmental gene regulation.


Assuntos
Quebra Cromossômica , Cromossomos/genética , Telômero/genética , Tetrahymena thermophila/genética , Animais , Núcleo Celular/genética , Instabilidade Cromossômica/genética , Bases de Dados Genéticas , Expressão Gênica/genética , Biblioteca Genômica , Células Germinativas/crescimento & desenvolvimento , Tetrahymena thermophila/crescimento & desenvolvimento
8.
Biol Open ; 5(12): 1790-1798, 2016 Dec 15.
Artigo em Inglês | MEDLINE | ID: mdl-27793833

RESUMO

The p68 DEAD box helicases comprise a widely conserved protein family involved in a large range of biological processes including transcription, splicing and translation. The genome of the ciliate Tetrahymena thermophile encodes two p68-like helicases, Drh1p and Lia2p. We show that DRH1 is essential for growth and completion of development. In growing cells, Drh1p is excluded from the nucleus and accumulates near cortical basal bodies. In contrast, during sexual reproduction, this protein localizes to meiotic micronuclei, initially in punctate foci in regions where centromeres and telomeres are known to reside and later in post-zygotic differentiating somatic macronuclei. Differentiation of the macronuclear genome involves extensive DNA rearrangements including fragmentation of the five pairs of germline-derived chromosomes into 180 chromosomal sub-fragments that are stabilized by de novo telomere deletion. In addition, thousands of internal eliminated sequences (IESs) are excised from loci dispersed throughout the genome. Strains with DRH1 deleted from the germline nuclei, which do not express the protein during post-zygotic development, fail to fragment the developing macronuclear chromosomes. IES excision still occurs in the absence of DRH1 zygotic expression; thus, Drh1p is the first protein found to be specifically required for chromosome breakage but not DNA elimination.

9.
J Cell Sci ; 129(5): 1046-58, 2016 Mar 01.
Artigo em Inglês | MEDLINE | ID: mdl-26769902

RESUMO

Bi-directional non-coding transcripts and their ∼29-nt small RNA products are known to guide DNA deletion in Tetrahymena, leading to the removal of one-third of the genome from developing somatic nuclei. Using an antibody specific for long double-stranded RNAs (dsRNAs), we determined the dynamic subcellular distributions of these RNAs. Conjugation-specific dsRNAs were found and show sequential appearances in parental germline, parental somatic nuclei and finally in new somatic nuclei of progeny. The dsRNAs in germline nuclei and new somatic nuclei are likely transcribed from the sequences destined for deletion; however, the dsRNAs in parental somatic nuclei are unexpected, and PCR analyses suggested that they were transcribed in this nucleus. Deficiency in the RNA interference (RNAi) pathway led to abnormal aggregations of dsRNA in both the parental and new somatic nuclei, whereas accumulation of dsRNAs in the germline nuclei was only seen in the Dicer-like gene mutant. In addition, RNAi mutants displayed an early loss of dsRNAs from developing somatic nuclei. Thus, long dsRNAs are made in multiple nuclear compartments and some are linked to small RNA production whereas others might participate in their regulations.


Assuntos
Núcleo Celular/fisiologia , RNA de Cadeia Dupla/metabolismo , RNA de Protozoário/metabolismo , Rearranjo Gênico , Genoma de Protozoário , Heterocromatina/metabolismo , Proteínas de Protozoários/genética , Proteínas de Protozoários/metabolismo , Transporte de RNA , RNA de Cadeia Dupla/genética , RNA de Protozoário/genética , Tetrahymena
10.
Genes Dev ; 30(24): 2724-2736, 2016 12 15.
Artigo em Inglês | MEDLINE | ID: mdl-28087716

RESUMO

Ciliated protozoans perform extreme forms of programmed somatic DNA rearrangement during development. The model ciliate Tetrahymena thermophila removes 34% of its germline micronuclear genome from somatic macronuclei by excising thousands of internal eliminated sequences (IESs), a process that shares features with transposon excision. Indeed, piggyBac transposon-derived genes are necessary for genome-wide IES excision in both Tetrahymena (TPB2 [Tetrahymena piggyBac-like 2] and LIA5) and Paramecium tetraurelia (PiggyMac). T. thermophila has at least three other piggyBac-derived genes: TPB1, TPB6, and TPB7 Here, we show that TPB1 and TPB6 excise a small, distinct set of 12 unusual IESs that disrupt exons. TPB1-deficient cells complete mating, but their progeny exhibit slow growth, giant vacuoles, and osmotic shock sensitivity due to retention of an IES in the vacuolar gene DOP1 (Dopey domain-containing protein). Unlike most IESs, TPB1-dependent IESs have piggyBac-like terminal inverted motifs that are necessary for excision. Transposon-like excision mediated by TPB1 and TPB6 provides direct evidence for a transposon origin of not only IES excision machinery but also IESs themselves. Our study highlights a division of labor among ciliate piggyBac-derived genes, which carry out mutually exclusive categories of excision events mediated by either transposon-like features or RNA-directed heterochromatin.


Assuntos
Elementos de DNA Transponíveis/genética , Rearranjo Gênico/genética , Genes de Protozoários/genética , Genoma de Protozoário/genética , Proteínas de Protozoários/metabolismo , Tetrahymena thermophila/genética , Regulação da Expressão Gênica no Desenvolvimento , Técnicas de Inativação de Genes , Estágios do Ciclo de Vida , Proteínas de Protozoários/genética , Tetrahymena thermophila/crescimento & desenvolvimento , Vacúolos/genética
11.
Microbiol Spectr ; 2(6)2014 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-26104448

RESUMO

Ciliates are champions in programmed genome rearrangements. They carry out extensive restructuring during differentiation to drastically alter the complexity, relative copy number, and arrangement of sequences in the somatic genome. This chapter focuses on the model ciliate Tetrahymena, perhaps the simplest and best-understood ciliate studied. It summarizes past studies on various genome rearrangement processes and describes in detail the remarkable progress made in the past decade on the understanding of DNA deletion and other processes. The process occurs at thousands of specific sites to remove defined DNA segments that comprise roughly one-third of the genome including all transposons. Interestingly, this DNA rearranging process is a special form of RNA interference. It involves the production of double-stranded RNA and small RNA that guides the formation of heterochromatin. A domesticated piggyBac transposase is believed to cut off the marked chromatin, and the retained sequences are joined together through nonhomologous end-joining processes. Many of the proteins and DNA players involved have been analyzed and are described. This link provides possible explanations for the evolution, mechanism, and functional roles of the process. The article also discusses the interactions between parental and progeny somatic nuclei that affect the selection of sequences for deletion, and how the specific deletion boundaries are determined after heterochromatin marking.


Assuntos
DNA de Protozoário/genética , Rearranjo Gênico , Genes de Protozoários , Genoma de Protozoário , Deleção de Sequência , Tetrahymena/genética , Interferência de RNA , RNA não Traduzido/metabolismo , Transposases/metabolismo
12.
Mol Biol Cell ; 23(11): 2213-25, 2012 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-22513090

RESUMO

Programmed DNA rearrangements are important processes present in many organisms. In the ciliated protozoan Tetrahymena thermophila, DNA rearrangements occur during the sexual conjugation process and lead to the deletion of thousands of specific DNA segments and fragmentation of the chromosomes. In this study, we found that the Ku80 homologue, a conserved component of the nonhomologous end-joining process of DNA repair, was essential for these two processes. During conjugation, TKU80 was highly expressed and localized to the new macronucleus, where DNA rearrangements occur. Homokaryon TKU80-knockout mutants are unable to complete conjugation and produce progeny and are arrested at the two-micronuclei/two-macronuclei stage. Analysis of their DNA revealed failure to complete DNA deletion. However, the DNA-cutting step appeared to have occurred, as evidenced by the presence of circularized excised DNA. Moreover, chromosome breakage or de novo telomere addition was affected. The mutant appears to accumulate free DNA ends detectable by terminal deoxynucleotidyl transferase dUTP nick end labeling assays that led to the degradation of most DNA in the developing macronucleus. These findings suggest that Tku80p may serve an end-protective role after DNA cleavage has occurred. Unexpectedly, the large heterochromatin structures that normally associate with DNA rearrangements failed to form without TKU80. Together the results suggest multiple roles for Tku80p and indicate that a Ku-dependent DNA-repair pathway is involved in programmed DNA rearrangements in Tetrahymena.


Assuntos
Antígenos Nucleares/metabolismo , Quebras de DNA , Reparo do DNA/genética , Proteínas de Ligação a DNA/metabolismo , Rearranjo Gênico/genética , Proteínas de Protozoários/metabolismo , Tetrahymena thermophila/genética , Sequência de Bases , Quebra Cromossômica , Conjugação Genética , DNA de Protozoário/genética , Proteínas de Fluorescência Verde/metabolismo , Histonas/metabolismo , Autoantígeno Ku , Macronúcleo/metabolismo , Metilação , Modelos Biológicos , Proteínas Nucleares/metabolismo , Fosfoproteínas/metabolismo , Transporte Proteico , Proteínas Recombinantes de Fusão/metabolismo , Tetrahymena thermophila/citologia
13.
Eukaryot Cell ; 11(5): 601-14, 2012 May.
Artigo em Inglês | MEDLINE | ID: mdl-22427430

RESUMO

Histone H3K27me3 modification is an important regulator for development and gene expression. In Tetrahymena thermophila, the complex chromatin dynamics of H3K27me3 marks during nuclear development suggested that an H3K27me3 demethylase might exist. Here, we report an H3K27me3 demethylase homolog, JMJ1, in Tetrahymena. During conjugation, JMJ1 expression is upregulated and the protein is localized first in the parental macronucleus and then in the new macronucleus. In conjugating cells, knockdown of JMJ1 expression resulted in a severe reduction in the production of progeny, suggesting that JMJ1 is essential for Tetrahymena conjugation. Furthermore, knockdown of JMJ1 resulted in increased H3K27 trimethylation in the new macronucleus and reduced transcription of genes related to DNA elimination, while the DNA elimination process was also partially blocked. Knockdown of the H3K27 methyltransferase EZL2 but not that of EZL1 partially restored progeny production in JMJ1-knockdown cells and reduced abnormal H3K27me3 accumulation in the new macronucleus. Taken together, these results demonstrate a critical role for JMJ1 in regulating H3K27me3 during conjugation and the importance of JMJ1 in regulating gene expression in the new macronucleus but not in regulating the formation of heterochromatin associated with programmed DNA deletion.


Assuntos
Histonas/metabolismo , Histona Desmetilases com o Domínio Jumonji/metabolismo , Proteínas de Protozoários/metabolismo , Tetrahymena thermophila/metabolismo , Sequência de Aminoácidos , Western Blotting , Cloreto de Cádmio/farmacologia , Imunoprecipitação da Cromatina , Quebra Cromossômica , Biologia Computacional , Conjugação Genética , DNA de Protozoário/genética , DNA de Protozoário/metabolismo , Técnicas de Silenciamento de Genes , Heterocromatina/genética , Heterocromatina/metabolismo , Histona Metiltransferases , Histona-Lisina N-Metiltransferase/genética , Histona-Lisina N-Metiltransferase/metabolismo , Macronúcleo/enzimologia , Macronúcleo/genética , Macronúcleo/metabolismo , Metilação , Dados de Sequência Molecular , Proteínas Nucleares/genética , Proteínas Nucleares/metabolismo , Fosfoproteínas/genética , Fosfoproteínas/metabolismo , Filogenia , Proteínas de Protozoários/genética , Interferência de RNA , RNA Mensageiro/genética , RNA Mensageiro/metabolismo , RNA de Protozoário/genética , Alinhamento de Sequência , Homologia de Sequência de Aminoácidos , Tetrahymena thermophila/enzimologia , Tetrahymena thermophila/genética , Transcrição Gênica , Ativação Transcricional
14.
Eukaryot Cell ; 11(4): 494-506, 2012 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-22366125

RESUMO

Autophagy is an evolutionarily conserved mechanism for the degradation of cellular components, but its role in enucleation during differentiation has not been established. Tetrahymena thermophila is a unicellular eukaryote with two functionally distinct nuclei, the somatic (macro-) and the germ line (micro-) nuclei. These nuclei are produced during sexual reproduction (conjugation), which involves differentiation and selective degradation of several specific nuclei. To examine the role of autophagy in nuclear degradation, we studied the function of two ATG8 genes in Tetrahymena. Through fluorescent protein tagging, we found that both proteins are targeted to degrading nuclei at specific stages, with some enrichment on the nuclear periphery, suggesting the formation of autophagosomes surrounding these nuclei. In addition, ATG8 knockout mutant cells showed a pronounced delay in nuclear degradation without apparently preventing the completion of other developmental events. This evidence provided direct support for a critical role for autophagy in programmed nuclear degradation. The results also showed differential roles for two ATG8 genes, with ATG8-65 playing a more significant role in starvation than ATG8-2, although both are important in nuclear degradation.


Assuntos
Autofagia/genética , Macronúcleo/metabolismo , Micronúcleo Germinativo/metabolismo , Proteínas de Protozoários/fisiologia , Tetrahymena thermophila/fisiologia , Sequência de Aminoácidos , Sequência Conservada , DNA de Protozoário/metabolismo , Viabilidade Microbiana , Dados de Sequência Molecular , Transporte Proteico , Proteínas de Protozoários/metabolismo , Proteínas Recombinantes de Fusão/metabolismo , Reprodução , Tetrahymena thermophila/genética , Tetrahymena thermophila/metabolismo
15.
Annu Rev Genet ; 45: 227-46, 2011.
Artigo em Inglês | MEDLINE | ID: mdl-21910632

RESUMO

Ciliated protozoa extensively remodel their somatic genomes during nuclear development, fragmenting their chromosomes and removing large numbers of internal eliminated sequences (IESs). The sequences eliminated are unique and repetitive DNAs, including transposons. Recent studies have identified transposase proteins that appear to have been domesticated and are used by these cells to eliminate DNA not wanted in the somatic macronucleus. This DNA elimination process is guided by meiotically produced small RNAs, generated in the germline nucleus, that recognize homologous sequences leading to their removal. These scan RNAs are found in complexes with PIWI proteins. Before they search the developing genome for IESs to eliminate, they scan the parental somatic nucleus and are removed from the pool if they match homologous sequences in that previously reorganized genome. In Tetrahymena, the scan RNAs target heterochromatin modifications to mark IESs for elimination. This DNA elimination pathway in ciliates shares extensive similarity with piRNA-mediated silencing of metazoans and highlights the remarkable ability of homologous RNAs to shape developing genomes.


Assuntos
Cilióforos/genética , DNA de Protozoário/genética , Genoma de Protozoário , Núcleo Celular/genética , Núcleo Celular/metabolismo , Cromossomos/genética , Cromossomos/metabolismo , Cilióforos/metabolismo , Elementos de DNA Transponíveis , DNA de Protozoário/metabolismo , Epigênese Genética , Regulação da Expressão Gênica , Rearranjo Gênico , Heterocromatina/genética , Heterocromatina/metabolismo , RNA de Protozoário/genética , RNA de Protozoário/metabolismo , Transposases/metabolismo
16.
J Mol Evol ; 72(5-6): 510-20, 2011 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-21643829

RESUMO

Centromere-drive is a process where centromeres compete for transmission through asymmetric "female" meiosis for inclusion into the oocyte. In symmetric "male" meiosis, all meiotic products form viable germ cells. Therefore, the primary incentive for centromere-drive, a potential transmission bias, is believed to be missing from male meiosis. In this article, we consider whether male meiosis also bears the primary cost of centromere-drive. Because different taxa carry out different combinations of meiotic programs (symmetric + asymmetric, symmetric only, asymmetric only), it is possible to consider the evolutionary consequences of centromere-drive in the context of these differing systems. Groups with both types of meiosis have large, rapidly evolving centromeric regions, and their centromeric histones (CenH3s) have been shown to evolve under positive selection, suggesting roles as suppressors of centromere-drive. In contrast, taxa with only symmetric male meiosis have shown no evidence of positive selection in their centromeric histones. In this article, we present the first evolutionary analysis of centromeric histones in ciliated protozoans, a group that only undergoes asymmetric "female" meiosis. We find no evidence of positive selection acting on CNA1, the CenH3 of Tetrahymena species. Cytological observations of a panel of Tetrahymena species are consistent with dynamic karyotype evolution in this lineage. Our findings suggest that defects in male meiosis, and not mitosis or female meiosis, are the primary selective force behind centromere-drive suppression. Our study raises the possibility that taxa like ciliates, with only female meiosis, may therefore undergo unsuppressed centromere drive.


Assuntos
Centrômero/genética , Histonas/metabolismo , Meiose/genética , Tetrahymena/genética , Tetrahymena/metabolismo , Sequência de Aminoácidos , Animais , Evolução Molecular , Feminino , Regulação da Expressão Gênica , Genes de Protozoários , Masculino , Dados de Sequência Molecular , Filogenia , Transporte Proteico , Alinhamento de Sequência , Tetrahymena/classificação
18.
Mol Biol Cell ; 21(10): 1753-62, 2010 May 15.
Artigo em Inglês | MEDLINE | ID: mdl-20357003

RESUMO

Transposons comprise large fractions of eukaryotic genomes and provide genetic reservoirs for the evolution of new cellular functions. We identified TPB2, a homolog of the piggyBac transposase gene that is required for programmed DNA deletion in Tetrahymena. TPB2 was expressed exclusively during the time of DNA excision, and its encoded protein Tpb2p was localized in DNA elimination heterochromatin structures. Notably, silencing of TPB2 by RNAi disrupts the final assembly of these heterochromatin structures and prevents DNA deletion to occur. In vitro studies revealed that Tpb2p is an endonuclease that produces double-strand breaks with four-base 5' protruding ends, similar to the ends generated during DNA deletion. These findings suggest that Tpb2p plays a key role in the assembly of specialized DNA elimination chromatin architectures and is likely responsible for the DNA cleavage step of programmed DNA deletion.


Assuntos
Heterocromatina/metabolismo , Tetrahymena thermophila/genética , Tetrahymena thermophila/metabolismo , Transposases/metabolismo , Cromatina , DNA/genética , Clivagem do DNA , Eucariotos , Heterocromatina/genética , Deleção de Sequência , Transposases/genética
20.
Nat Rev Cancer ; 9(3): 216-24, 2009 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-19212324

RESUMO

The clinical importance of gene amplification in the diagnosis and treatment of cancer has been widely recognized, as it is often evident in advanced stages of diseases. However, our knowledge of the underlying mechanisms is still limited. Gene amplification is an essential process in several organisms including the ciliate Tetrahymena thermophila, in which the initiating mechanism has been well characterized. Lessons from such simple eukaryotes may provide useful information regarding how gene amplification occurs in tumour cells.


Assuntos
Amplificação de Genes , Genoma , Sequências Repetidas Invertidas , Neoplasias/genética , Animais , Evolução Biológica , Quebras de DNA de Cadeia Dupla , Humanos , Tetrahymena thermophila/genética
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