Deadenylase depletion protects inherited mRNAs in primordial germ cells

Development. 2014 Aug;141(16):3134-42. doi: 10.1242/dev.110395.

Abstract

A crucial event in animal development is the specification of primordial germ cells (PGCs), which become the stem cells that create sperm and eggs. How PGCs are created provides a valuable paradigm for understanding stem cells in general. We find that the PGCs of the sea urchin Strongylocentrotus purpuratus exhibit broad transcriptional repression, yet enrichment for a set of inherited mRNAs. Enrichment of several germline determinants in the PGCs requires the RNA-binding protein Nanos to target the transcript that encodes CNOT6, a deadenylase, for degradation in the PGCs, thereby creating a stable environment for RNA. Misexpression of CNOT6 in the PGCs results in their failure to retain Seawi transcripts and Vasa protein. Conversely, broad knockdown of CNOT6 expands the domain of Seawi RNA as well as exogenous reporters. Thus, Nanos-dependent spatially restricted CNOT6 differential expression is used to selectively localize germline RNAs to the PGCs. Our findings support a 'time capsule' model of germline determination, whereby the PGCs are insulated from differentiation by retaining the molecular characteristics of the totipotent egg and early embryo.

Keywords: CCR4; CNOT6; Germ line; Nanos; PGC; Pumilio; Sea urchin; Transcriptomics.

Publication types

  • Research Support, N.I.H., Extramural
  • Research Support, U.S. Gov't, Non-P.H.S.

MeSH terms

  • Animals
  • Base Sequence
  • Cell Differentiation
  • Cell Separation
  • Exoribonucleases / metabolism*
  • Flow Cytometry
  • Gene Expression Profiling
  • Gene Expression Regulation, Developmental*
  • Germ Cells / cytology*
  • Molecular Sequence Data
  • RNA, Messenger / metabolism
  • RNA-Binding Proteins / metabolism
  • Strongylocentrotus purpuratus / embryology*
  • Strongylocentrotus purpuratus / enzymology
  • Time Factors
  • Transcriptome

Substances

  • RNA, Messenger
  • RNA-Binding Proteins
  • Exoribonucleases