Genetic requirements for PIE-1 localization and inhibition of gene expression in the embryonic germ lineage of Caenorhabditis elegans.
In early Caenorhabditis elegans embryos, production of new mRNAs is inhibited in the germ lineage. This inhibition requires the germline factor PIE-1, and correlates with the absence in germline blastomeres of a phosphoepitope on RNA polymerase II (RNAPII-H5). We show that PIE-1 is uniformly distributed in oocytes and newly fertilized eggs, and becomes localized asymmetrically in the late one-cell stage. To begin to dissect the mechanisms required for PIE-1 localization and inhibition of RNAPII-H5 expression, we have examined the distribution of PIE-1 and RNAPII-H5 in maternal-effect mutants that disrupt embryonic development. We find that mutants that disrupt the asymmetric divisions of germline blastomeres mislocalize PIE-1, and activate RNAPII-H5 expression in the germ lineage. In contrast, mutants that alter somatic cell identities do not affect PIE-1 localization or RNAPII-H5 expression. Our observations suggest that PIE-1 represses mRNA transcription in each germline blastomere in a concentration-dependent manner. We also show that in wild-type, and in mutants where PIE-1 is mislocalized, the cellular and subcellular distribution of PIE-1 remarkably parallels that of the P granules, suggesting that the localizations of these two germline components are coordinately regulated.
School of Medicine, Johns Hopkins University, Baltimore, Maryland, 21205-2185, USA.
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Tenenhaus
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Schubert
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Seydoux
D000818:Animals; D000911:Antibodies, Monoclonal / Q000378:metabolism; D017173:Caenorhabditis elegans / Q000196:embryology*; D029742:Caenorhabditis elegans Proteins; D003594:Cytoplasmic Granules / Q000378:metabolism; D004247:DNA / Q000032:analysis; D005455:Fluorescent Antibody Technique; D018507:Gene Expression Regulation, Developmental / Q000235:genetics*; D005854:Germ Cells / Q000378:metabolism*; D006579:Heterozygote; D017403:In Situ Hybridization; D009154:Mutation / Q000235:genetics; D009687:Nuclear Proteins / Q000276:immunology / Q000378:metabolism*; D009865:Oocytes / Q000737:chemistry; D010641:Phenotype; D012319:RNA Polymerase II / Q000378:metabolism; D012333:RNA, Messenger / Q000378:metabolism; D015533:Transcriptional Activation / Q000502:physiology
D000911:Antibodies, Monoclonal; D029742:Caenorhabditis elegans Proteins; D009687:Nuclear Proteins; D012333:RNA, Messenger; C100950:pie-1 protein, C elegans; D004247:DNA; D012319:RNA Polymerase II