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Dynamic regulatory phosphorylation of mouse CDK2 occurs during meiotic prophase I

During prophase I of meiosis, DNA double-strand breaks form throughout the genome, with a subset repairing as crossover events, enabling the accurate segregation of homologous chromosomes during the first meiotic division. The mechanism by which DSBs become selected to repair as crossovers is unknow...

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Autores principales: Bradley, Rachel A., Wolff, Ian D., Cohen, Paula E., Gray, Stephen
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Cold Spring Harbor Laboratory 2023
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10402020/
https://www.ncbi.nlm.nih.gov/pubmed/37546989
http://dx.doi.org/10.1101/2023.07.24.550435
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author Bradley, Rachel A.
Wolff, Ian D.
Cohen, Paula E.
Gray, Stephen
author_facet Bradley, Rachel A.
Wolff, Ian D.
Cohen, Paula E.
Gray, Stephen
author_sort Bradley, Rachel A.
collection PubMed
description During prophase I of meiosis, DNA double-strand breaks form throughout the genome, with a subset repairing as crossover events, enabling the accurate segregation of homologous chromosomes during the first meiotic division. The mechanism by which DSBs become selected to repair as crossovers is unknown, although the crossover positioning and levels in each cell indicate it is a highly regulated process. One of the proteins that localises to crossover sites is the serine/threonine cyclin-dependent kinase CDK2. Regulation of CDK2 occurs via phosphorylation at tyrosine 15 (Y15) and threonine 160 (T160) inhibiting and activating the kinase, respectively. In this study we use a combination of immunofluorescence staining on spread spermatocytes and fixed testis sections, and STA-PUT gravitational sedimentation to isolate cells at different developmental stages to further investigate the temporal phospho regulation of CDK2 during prophase I. Western blotting reveals differential levels of the two CDK2 isoforms (CDK2(33kDa) and CDK2(39kDa)) throughout prophase I, with inhibitory phosphorylation of CDK2 at Y15 occurring early in prophase I, localising to telomeres and diminishing as cells enter pachynema. Conversely, the activatory phosphorylation on T160 occurs later, specifically the CDK2(33kDa) isoform, and T160 signal is detected in spermatogonia and pachytene spermatocytes, where it co-localises with the Class I crossover protein MLH3. Taken together, our data reveals intricate control of CDK2 both with regards to levels of the two CDK2 isoforms, and differential regulation via inhibitory and activatory phosphorylation.
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spelling pubmed-104020202023-08-05 Dynamic regulatory phosphorylation of mouse CDK2 occurs during meiotic prophase I Bradley, Rachel A. Wolff, Ian D. Cohen, Paula E. Gray, Stephen bioRxiv Article During prophase I of meiosis, DNA double-strand breaks form throughout the genome, with a subset repairing as crossover events, enabling the accurate segregation of homologous chromosomes during the first meiotic division. The mechanism by which DSBs become selected to repair as crossovers is unknown, although the crossover positioning and levels in each cell indicate it is a highly regulated process. One of the proteins that localises to crossover sites is the serine/threonine cyclin-dependent kinase CDK2. Regulation of CDK2 occurs via phosphorylation at tyrosine 15 (Y15) and threonine 160 (T160) inhibiting and activating the kinase, respectively. In this study we use a combination of immunofluorescence staining on spread spermatocytes and fixed testis sections, and STA-PUT gravitational sedimentation to isolate cells at different developmental stages to further investigate the temporal phospho regulation of CDK2 during prophase I. Western blotting reveals differential levels of the two CDK2 isoforms (CDK2(33kDa) and CDK2(39kDa)) throughout prophase I, with inhibitory phosphorylation of CDK2 at Y15 occurring early in prophase I, localising to telomeres and diminishing as cells enter pachynema. Conversely, the activatory phosphorylation on T160 occurs later, specifically the CDK2(33kDa) isoform, and T160 signal is detected in spermatogonia and pachytene spermatocytes, where it co-localises with the Class I crossover protein MLH3. Taken together, our data reveals intricate control of CDK2 both with regards to levels of the two CDK2 isoforms, and differential regulation via inhibitory and activatory phosphorylation. Cold Spring Harbor Laboratory 2023-07-24 /pmc/articles/PMC10402020/ /pubmed/37546989 http://dx.doi.org/10.1101/2023.07.24.550435 Text en https://creativecommons.org/licenses/by-nc-nd/4.0/This work is licensed under a Creative Commons Attribution-NonCommercial-NoDerivatives 4.0 International License (https://creativecommons.org/licenses/by-nc-nd/4.0/) , which allows reusers to copy and distribute the material in any medium or format in unadapted form only, for noncommercial purposes only, and only so long as attribution is given to the creator.
spellingShingle Article
Bradley, Rachel A.
Wolff, Ian D.
Cohen, Paula E.
Gray, Stephen
Dynamic regulatory phosphorylation of mouse CDK2 occurs during meiotic prophase I
title Dynamic regulatory phosphorylation of mouse CDK2 occurs during meiotic prophase I
title_full Dynamic regulatory phosphorylation of mouse CDK2 occurs during meiotic prophase I
title_fullStr Dynamic regulatory phosphorylation of mouse CDK2 occurs during meiotic prophase I
title_full_unstemmed Dynamic regulatory phosphorylation of mouse CDK2 occurs during meiotic prophase I
title_short Dynamic regulatory phosphorylation of mouse CDK2 occurs during meiotic prophase I
title_sort dynamic regulatory phosphorylation of mouse cdk2 occurs during meiotic prophase i
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10402020/
https://www.ncbi.nlm.nih.gov/pubmed/37546989
http://dx.doi.org/10.1101/2023.07.24.550435
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