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1.
PLoS Genet ; 14(5): e1007388, 2018 05.
Artigo em Inglês | MEDLINE | ID: mdl-29813053

RESUMO

It is widely accepted in eukaryotes that the cleavage furrow only initiates after mitosis completion. In fission yeast, cytokinesis requires the synthesis of a septum tightly coupled to cleavage furrow ingression. The current cytokinesis model establishes that simultaneous septation and furrow ingression only initiate after spindle breakage and mitosis exit. Thus, this model considers that although Cdk1 is inactivated at early-anaphase, septation onset requires the long elapsed time until mitosis completion and full activation of the Hippo-like SIN pathway. Here, we studied the precise timing of septation onset regarding mitosis by exploiting both the septum-specific detection with the fluorochrome calcofluor and the high-resolution electron microscopy during anaphase and telophase. Contrarily to the existing model, we found that both septum and cleavage furrow start to ingress at early anaphase B, long before spindle breakage, with a slow ingression rate during anaphase B, and greatly increasing after telophase onset. This shows that mitosis and cleavage furrow ingression are not concatenated but simultaneous events in fission yeast. We found that the timing of septation during early anaphase correlates with the cell size and is regulated by the corresponding levels of SIN Etd1 and Rho1. Cdk1 inactivation was directly required for timely septation in early anaphase. Strikingly the reduced SIN activity present after Cdk1 loss was enough to trigger septation by immediately inducing the medial recruitment of the SIN kinase complex Sid2-Mob1. On the other hand, septation onset did not depend on the SIN asymmetry establishment, which is considered a hallmark for SIN activation. These results recalibrate the timing of key cytokinetic events in fission yeast; and unveil a size-dependent control mechanism that synchronizes simultaneous nuclei separation with septum and cleavage furrow ingression to safeguard the proper chromosome segregation during cell division.


Assuntos
Anáfase/fisiologia , Proteínas de Ciclo Celular/fisiologia , Citocinese/fisiologia , Proteínas de Schizosaccharomyces pombe/fisiologia , Schizosaccharomyces/fisiologia , Fuso Acromático/fisiologia , Benzenossulfonatos/química , Proteína Quinase CDC2/fisiologia , Núcleo Celular/fisiologia , Microscopia Eletrônica de Transmissão , Microscopia de Fluorescência/métodos , Proteínas Quinases/fisiologia , Schizosaccharomyces/ultraestrutura , Fuso Acromático/ultraestrutura , Telófase/fisiologia , Fatores de Tempo , Proteínas rho de Ligação ao GTP/fisiologia
2.
Int J Food Microbiol ; 238: 146-152, 2016 Dec 05.
Artigo em Inglês | MEDLINE | ID: mdl-27620826

RESUMO

The aim of this study was to assess the species and the genetic diversity of the staphylococci population in raw milk from healthy goats. Isolates representative of all genotypes were screened for their potential pathogenicity by the occurrence of some relevant safety-related properties, such as antibiotic resistance, presence of virulence factor genes, biofilm formation ability and biogenic amine production. A total of 314 staphylococci were isolated, and randomly amplified polymorphic DNA-PCR analysis displayed 48 genotypes. Isolates were identified as belonging to S. epidermidis (87.5%), S. caprae (6.2%), S. aureus (4.2%) and S. simulans (2.1%) species. The antibiotic resistance varied strongly with strains, with S. epidermidis and S. aureus strains showing resistance to more number of antibiotics. A high occurrence of strains harbouring hemolysin genes was also found in both species. On the contrary, none of the strains assayed harboured enterotoxin or amino acid decarboxylase genes, and, although a moderate or high biofilm formation was observed in 29% of the strains, they did not harbour icaA or icaD genes. This study gives a first and extensive picture of safety-related properties within Staphylococcus species isolated from milk of healthy goats, displaying that these species can act as a reservoir for spreading genes related to safety.


Assuntos
Farmacorresistência Bacteriana , Microbiologia de Alimentos , Leite/microbiologia , Staphylococcus aureus/genética , Staphylococcus epidermidis/genética , Aminas/química , Animais , Antibacterianos/farmacologia , Biofilmes , Resistência Microbiana a Medicamentos/genética , Enterotoxinas/genética , Genótipo , Cabras , Fenótipo , Infecções Estafilocócicas/veterinária , Staphylococcus/genética , Fatores de Virulência/genética
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