TY - JOUR
T1 - Phosphorylation of HORMA-domain protein HTP-3 at Serine 285 is dispensable for crossover formation
AU - Das, Debabrata
AU - Trivedi, Shalini
AU - Blazıcková, Jitka
AU - Arur, Swathi
AU - Silva, Nicola
N1 - Funding Information:
The authors are grateful to Yumi Kim for sharing the panHTP-3 antibody and the AID::cdk-1 and cdk-2::AID::3xFLAG lines prior to publication, to Enrique Martinez-Perez for the chk-2::AID strain, and to Verena Jantsch and Angela Graf for the chk-1RNAi clone and the invaluable help with the microinjections. Some strains were provided by the CGC, which is funded by the NIH Office of Research Infrastructure Programs (P40 OD010440). Proteomics analyses were performed by the Mass Spectrometry Facility at Max Perutz Labs using the VBCF instrument pool, with special help from T. Gossenreiter and M. Hartl. They acknowledge the core facility CELLIM supported by the Czech-BioImaging large RI project (LM2018129 funded by MEYS CR) for their support with obtaining scientific data presented in this article.
Funding Information:
Research in NS laboratory is funded by GACR≤ (GA20–08819S) and a Start-Up grant from the Department of Biology of Masaryk University. DD is funded through Odyssey Postdoctoral Fellowship (supported by the Kimberly-Clark Foundation), UT MD Anderson Cancer Center. Parts of research reported in this publication were supported by the National Institute of Child Health and Development of the National Institute of Health under award number R01 HD101269 to SA.
Publisher Copyright:
© The Author(s) 2022.
PY - 2022/5
Y1 - 2022/5
N2 - Generation of functional gametes is accomplished through a multilayered and finely orchestrated succession of events during meiotic progression. In the Caenorhabditis elegans germline, the HORMA-domain-containing protein HTP-3 plays pivotal roles for the establishment of chromosome axes and the efficient induction of programmed DNA double-strand breaks, both of which are crucial for crossover formation. Double-strand breaks allow for accurate chromosome segregation during the first meiotic division and therefore are an essential requirement for the production of healthy gametes. Phosphorylation-dependent regulation of HORMAD protein plays important roles in controlling meiotic chromosome behavior. Here, we document a phospho-site in HTP-3 at Serine 285 that is constitutively phosphorylated during meiotic prophase I. pHTP-3S285 localization overlaps with panHTP-3 except in nuclei undergoing physiological apoptosis, in which pHTP-3 is absent. Surprisingly, we observed that phosphorylation of HTP-3 at S285 is independent of the canonical kinases that control meiotic progression in nematodes. During meiosis, the htp-3(S285A) mutant displays accelerated RAD-51 turnover, but no other meiotic abnormalities. Altogether, these data indicate that the Ser285 phosphorylation is independent of canonical meiotic protein kinases and does not regulate HTP-3-dependent meiotic processes. We propose a model wherein phosphorylation of HTP-3 occurs through noncanonical or redundant meiotic kinases and/or is likely redundant with additional phospho-sites for function in vivo.
AB - Generation of functional gametes is accomplished through a multilayered and finely orchestrated succession of events during meiotic progression. In the Caenorhabditis elegans germline, the HORMA-domain-containing protein HTP-3 plays pivotal roles for the establishment of chromosome axes and the efficient induction of programmed DNA double-strand breaks, both of which are crucial for crossover formation. Double-strand breaks allow for accurate chromosome segregation during the first meiotic division and therefore are an essential requirement for the production of healthy gametes. Phosphorylation-dependent regulation of HORMAD protein plays important roles in controlling meiotic chromosome behavior. Here, we document a phospho-site in HTP-3 at Serine 285 that is constitutively phosphorylated during meiotic prophase I. pHTP-3S285 localization overlaps with panHTP-3 except in nuclei undergoing physiological apoptosis, in which pHTP-3 is absent. Surprisingly, we observed that phosphorylation of HTP-3 at S285 is independent of the canonical kinases that control meiotic progression in nematodes. During meiosis, the htp-3(S285A) mutant displays accelerated RAD-51 turnover, but no other meiotic abnormalities. Altogether, these data indicate that the Ser285 phosphorylation is independent of canonical meiotic protein kinases and does not regulate HTP-3-dependent meiotic processes. We propose a model wherein phosphorylation of HTP-3 occurs through noncanonical or redundant meiotic kinases and/or is likely redundant with additional phospho-sites for function in vivo.
KW - Caenorhabditis elegans meiosis
KW - HORMA-domain proteins
KW - HTP-3
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U2 - 10.1093/g3journal/jkac079
DO - 10.1093/g3journal/jkac079
M3 - Article
C2 - 35389463
AN - SCOPUS:85129997841
SN - 2160-1836
VL - 12
JO - G3: Genes, Genomes, Genetics
JF - G3: Genes, Genomes, Genetics
IS - 5
M1 - jkac079
ER -