RRC ID 56185
著者 Kocsisova Z, Kornfeld K, Schedl T.
タイトル Cell cycle accumulation of the proliferating cell nuclear antigen PCN-1 transitions from continuous in the adult germline to intermittent in the early embryo of C. elegans.
ジャーナル BMC Dev Biol
Abstract BACKGROUND:The proliferating cell nuclear antigen (PCNA or PCN-1 in C. elegans), an essential processivity factor for DNA polymerase δ, has been widely used as a marker of S-phase. In C. elegans early embryos, PCN-1 accumulation is cyclic, localizing to the nucleus during S-phase and the cytoplasm during the rest of the cell cycle. The C. elegans larval and adult germline is an important model systems for studying cell cycle regulation, and it was observed that the cell cycle regulator cyclin E (CYE-1 in C. elegans) displays a non-cyclic, continuous accumulation pattern in this tissue. The accumulation pattern of PCN-1 has not been well defined in the larval and adult germline, and the objective of this study was to determine if the accumulation pattern is cyclic, as in other cells and organisms, or continuous, similar to cyclin E.
RESULTS:To study the larval and adult germline accumulation of PCN-1 expressed from its native locus, we used CRISPR/Cas9 technology to engineer a novel allele of pcn-1 that encodes an epitope-tagged protein. S-phase nuclei were labeled using EdU nucleotide incorporation, and FLAG::PCN-1 was detected by antibody staining. All progenitor zone nuclei, including those that were not in S-phase (as they were negative for EdU staining) showed PCN-1 accumulation, indicating that PCN-1 accumulated during all cell cycle phases in the germline progenitor zone. The same result was observed with a GFP::PCN-1 fusion protein expressed from a transgene. pcn-1 loss-of-function mutations were analyzed, and pcn-1 was necessary for robust fertility and embryonic development.
CONCLUSIONS:In the C. elegans early embryo as well as other organisms, PCN-1 accumulates in nuclei only during S-phase. By contrast, in the progenitor zone of the germline of C. elegans, PCN-1 accumulated in nuclei during all cell cycle stages. This pattern is similar to accumulation pattern of cyclin E. These observations support the model that mitotic cell cycle regulation in the germline stem and progenitor cells is distinct from somatic cells, as it does not heavily rely on cyclic accumulation of classic cell cycle proteins.
巻・号 18(1)
ページ 12
公開日 2018-5-30
DOI 10.1186/s12861-018-0171-7
PII 10.1186/s12861-018-0171-7
PMID 29848313
PMC PMC5977546
MeSH Animals Caenorhabditis elegans / cytology Caenorhabditis elegans / embryology* Caenorhabditis elegans / metabolism* Caenorhabditis elegans Proteins / metabolism* Cell Cycle* Cell Nucleus / metabolism Embryo, Nonmammalian / cytology* Embryo, Nonmammalian / metabolism* Germ Cells / metabolism* Models, Biological Proliferating Cell Nuclear Antigen / metabolism* Stem Cells / cytology Stem Cells / metabolism
IF 2.368
引用数 2
リソース情報
線虫 tm3241