Post-transcriptional regulation of meiotic genes by a nuclear RNA silencing complex.
RNA is a central component of gene-silencing pathways that regulate diverse cellular processes. In the fission yeast Schizosaccharomyces pombe, an RNA-based mechanism represses meiotic gene expression during vegetative growth. This pathway depends on the zinc finger protein Red1, which is required to degrade meiotic mRNAs as well as to target histone H3 lysine 9 (H3K9) methylation, a repressive chromatin mark, to a subset of meiotic genes. However, the mechanism of Red1 function is unknown. Here we use affinity purification and mass spectrometry to identify a Red1-containing nuclear RNA silencing (NURS) complex. In addition to Red1, this complex includes the Mtl1, Red5, Ars2, Rmn1, and Iss10 proteins and associates with several other complexes that are involved in either signaling or mediating RNA silencing. By analyzing the effects of gene knockouts and inducible knockdown alleles, we show that NURS subunits regulate RNA degradation and H3K9 methylation at meiotic genes. We also identify roles for individual NURS subunits in interactions with Mmi1, an RNA-binding protein that marks meiotic RNAs for destruction, and the nuclear exosome RNA degradation complex. Finally, we show that the levels of H3K9 methylation at meiotic genes are not sufficient to restrict RNA polymerase II access or repress gene expression during vegetative growth. Our results demonstrate that Red1 partners with other proteins to silence meiotic gene expression at the post-transcriptional level. Conservation of a NURS-like complex in human cells suggests that this pathway plays an ancient and fundamental role in RNA silencing.
Howard Hughes Medical Institute, Harvard Medical School, Boston, Massachusetts 02115, USA Department of Cell Biology, Harvard Medical School, Boston, Massachusetts 02115, USA.
Emily D
ED
Egan
Department of Cell Biology, Harvard Medical School, Boston, Massachusetts 02115, USA.
Craig R
CR
Braun
Department of Cell Biology, Harvard Medical School, Boston, Massachusetts 02115, USA.
Steven P
SP
Gygi
Howard Hughes Medical Institute, Harvard Medical School, Boston, Massachusetts 02115, USA Department of Cell Biology, Harvard Medical School, Boston, Massachusetts 02115, USA.
Danesh
D
Moazed
D002352:Carrier Proteins / Q000235:genetics; D002843:Chromatin / Q000235:genetics; D055354:Exosomes / Q000235:genetics; D015966:Gene Expression Regulation, Fungal / Q000235:genetics; D006657:Histones / Q000235:genetics; D008540:Meiosis / Q000235:genetics*; D008745:Methylation; D034622:RNA Interference / Q000502:physiology*; D012319:RNA Polymerase II / Q000235:genetics; D020871:RNA Stability / Q000235:genetics; D012333:RNA, Messenger / Q000235:genetics; D015138:RNA, Nuclear / Q000235:genetics*; D029702:Schizosaccharomyces pombe Proteins / Q000235:genetics; D039221:mRNA Cleavage and Polyadenylation Factors / Q000235:genetics
D002352:Carrier Proteins; D002843:Chromatin; D006657:Histones; C512176:Mmi1 protein, S pombe; D012333:RNA, Messenger; D015138:RNA, Nuclear; C586176:Red1 protein, S pombe; D029702:Schizosaccharomyces pombe Proteins; D039221:mRNA Cleavage and Polyadenylation Factors; D012319:RNA Polymerase II
H3K9 methylation; Red1; fission yeast; mRNA degradation; meiosis; nuclear bodies