RNAi-mediated depletion of the 15 KH domain protein, vigilin, induces death of dividing and non-dividing human cells but does not initially inhibit protein synthesis.
Vigilin/Scp160p/DDP1 is a ubiquitous and highly conserved protein containing 15 related, but non-identical, K-homology (KH) nucleic acid binding domains. While its precise function remains unknown, proposed roles for vigilin include chromosome partitioning at mitosis, facilitating translation and tRNA transport, and control of mRNA metabolism, including estrogen-mediated stabilization of vitellogenin mRNA. To probe sites of vigilin action in vertebrate cells, we performed nucleic acid binding and RNA interference studies. Consistent with a potential role in chromosome partitioning, human vigilin exhibits a higher affinity for Drosophila dodecasatellite single-stranded DNA than for vitellogenin mRNA 3'-UTR. Direct observation and flow cytometry in non-mitotic, serum-starved, HeLa cells showed that RNAi-mediated vigilin knockdown is rapidly lethal, indicating an essential function for vigilin distinct from its proposed role in chromosome partitioning. Pulse labeling experiments revealed that rates of protein synthesis and degradation are unaffected by the several fold reduction in vigilin levels early in siRNA knockdown indicating that vigilin is not a global regulator of translation. These data show that vigilin is an essential protein in human cells, support the view that vigilin's most essential functions are neither chromosome partitioning nor control of translation, and are consistent with vigilin playing a critical role in cytoplasmic mRNA metabolism.
D020413:3' Untranslated Regions / Q000378:metabolism; D000818:Animals; D002352:Carrier Proteins; D016923:Cell Death; D002455:Cell Division; D002460:Cell Line; D002470:Cell Survival; D004276:DNA, Satellite / Q000378:metabolism; D004277:DNA, Single-Stranded / Q000378:metabolism; D004330:Drosophila / Q000235:genetics; D006367:HeLa Cells; D006801:Humans; D007700:Kinetics; D014176:Protein Biosynthesis; D017434:Protein Structure, Tertiary; D011506:Proteins / Q000378:metabolism; D034622:RNA Interference; D016601:RNA-Binding Proteins / Q000037:antagonists & inhibitors / Q000737:chemistry / Q000235:genetics / Q000502:physiology*; D014819:Vitellogenins / Q000235:genetics
D020413:3' Untranslated Regions; D002352:Carrier Proteins; D004276:DNA, Satellite; D004277:DNA, Single-Stranded; D011506:Proteins; D016601:RNA-Binding Proteins; D014819:Vitellogenins; C075197:high density lipoprotein binding protein