title: Deadenylase depletion protects inherited mRNAs in primordial germ cells.
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.
authors.affiliation: Department of Molecular, Cellular Biology and Biochemistry, Brown University, Providence, RI 02912, USA.
authors.forename: S Zachary
authors.initials: SZ
authors.lastname: Swartz
authors.affiliation: Department of Molecular, Cellular Biology and Biochemistry, Brown University, Providence, RI 02912, USA.
authors.forename: Adrian M
authors.initials: AM
authors.lastname: Reich
authors.affiliation: Department of Molecular, Cellular Biology and Biochemistry, Brown University, Providence, RI 02912, USA.
authors.forename: Nathalie
authors.initials: N
authors.lastname: Oulhen
authors.affiliation: Helicos BioSciences Corporation, One Kendall Square, Building 700, Cambridge, MA 02139, USA.
authors.forename: Tal
authors.initials: T
authors.lastname: Raz
authors.affiliation: Helicos BioSciences Corporation, One Kendall Square, Building 700, Cambridge, MA 02139, USA.
authors.forename: Patrice M
authors.initials: PM
authors.lastname: Milos
authors.affiliation: Marine Biology Research Division, Scripps Institution of Oceanography, University of California at San Diego, 8750 Biological Grade Road, La Jolla, CA 92037, USA.
authors.forename: Joseph P
authors.initials: JP
authors.lastname: Campanale
authors.affiliation: Marine Biology Research Division, Scripps Institution of Oceanography, University of California at San Diego, 8750 Biological Grade Road, La Jolla, CA 92037, USA.
authors.forename: Amro
authors.initials: A
authors.lastname: Hamdoun
authors.affiliation: Department of Molecular, Cellular Biology and Biochemistry, Brown University, Providence, RI 02912, USA rhet@brown.edu.
authors.forename: Gary M
authors.initials: GM
authors.lastname: Wessel
mesh_terms: D000818:Animals; D001483:Base Sequence; D002454:Cell Differentiation; D002469:Cell Separation; D005095:Exoribonucleases / Q000378:metabolism*; D005434:Flow Cytometry; D020869:Gene Expression Profiling; D018507:Gene Expression Regulation, Developmental; D005854:Germ Cells / Q000166:cytology*; D008969:Molecular Sequence Data; D012333:RNA, Messenger / Q000378:metabolism; D016601:RNA-Binding Proteins / Q000378:metabolism; D047270:Strongylocentrotus purpuratus / Q000196:embryology* / Q000201:enzymology; D013997:Time Factors; D059467:Transcriptome
chemical_list: D012333:RNA, Messenger; D016601:RNA-Binding Proteins; D005095:Exoribonucleases
keywords: CCR4; CNOT6; Germ line; Nanos; PGC; Pumilio; Sea urchin; Transcriptomics
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