CDK phosphorylation of SLD-2 is required for replication initiation and germline development in C. elegans

J Cell Biol. 2014 Feb 17;204(4):507-22. doi: 10.1083/jcb.201310083.

Abstract

Cyclin-dependent kinase (CDK) plays a vital role in proliferation control across eukaryotes. Despite this, how CDK mediates cell cycle and developmental transitions in metazoa is poorly understood. In this paper, we identify orthologues of Sld2, a CDK target that is important for DNA replication in yeast, and characterize SLD-2 in the nematode worm Caenorhabditis elegans. We demonstrate that SLD-2 is required for replication initiation and the nuclear retention of a critical component of the replicative helicase CDC-45 in embryos. SLD-2 is a CDK target in vivo, and phosphorylation regulates the interaction with another replication factor, MUS-101. By mutation of the CDK sites in sld-2, we show that CDK phosphorylation of SLD-2 is essential in C. elegans. Finally, using a phosphomimicking sld-2 mutant, we demonstrate that timely CDK phosphorylation of SLD-2 is an important control mechanism to allow normal proliferation in the germline. These results determine an essential function of CDK in metazoa and identify a developmental role for regulated SLD-2 phosphorylation.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Amino Acid Sequence
  • Animals
  • Blotting, Western
  • Caenorhabditis elegans / genetics*
  • Caenorhabditis elegans / growth & development
  • Caenorhabditis elegans / metabolism
  • Caenorhabditis elegans Proteins / genetics
  • Caenorhabditis elegans Proteins / metabolism*
  • Cell Proliferation
  • Cyclin-Dependent Kinases / genetics
  • Cyclin-Dependent Kinases / metabolism*
  • DNA Replication*
  • Embryo, Nonmammalian / cytology
  • Embryo, Nonmammalian / metabolism*
  • Fluorescent Antibody Technique
  • Gene Expression Regulation, Developmental
  • Germ Cells / physiology*
  • Molecular Sequence Data
  • Mutation / genetics
  • Phosphorylation
  • Sequence Homology, Amino Acid
  • Transgenes / physiology

Substances

  • Caenorhabditis elegans Proteins
  • Cyclin-Dependent Kinases