The human DEVH-box protein Ski2w from the HLA is localized in nucleoli and ribosomes.
The human helicase gene SKI2W is located between RD and RP1 in the class III region of the major histocompatibility complex. Transcripts of SKI2W are detectable in RNA samples isolated from multiple tissues. The protein product Ski2w shares striking amino acid sequence similarities to the yeast antiviral protein Ski2p that controls the translation of mRNAs, probably based on the mRNA structural integrity. Whether this translational regulation mechanism for cellular and viral RNAs exists in mammals is under investigation. Antisera against human Ski2w were generated using fusion proteins produced in bacteria or insect cells. Western blot analysis showed that the endogenous Ski2w protein is approximately 140 kDa in size and is enriched in polysomal fractions of cytoplasmic extracts from HeLa cells. Ribosomal profile studies revealed that Ski2w distributed throughout the entire sucrose gradient in the presence of Mg2+, but co-sedimented with the 18S rRNA-containing 40S subunit and the small ribosomal subunit protein S27a in the presence of EDTA. The co-sedimentation of Ski2w with the 40S subunit is not affected by RNase A treatment of the cell extract, or the addition of KCl to 0.5 M, suggesting that Ski2w is associated with the 40S ribosomal subunit. Indirect immunofluorescence experiments showed that human Ski2w is localized in the nucleoli and in the cytoplasm. In essence, human Ski2w is present at the sites of ribosome biogenesis and protein synthesis.
Children's Hospital Research Foundation, Molecular, Cellular and Developmental Biology Program, Department of Pediatrics, The Ohio StateUniversity, Columbus, OH 43205-2696, USA.
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D000595:Amino Acid Sequence; D000818:Animals; D016466:CHO Cells; D002451:Cell Compartmentation; D002466:Cell Nucleolus / Q000737:chemistry*; D006224:Cricetinae; D004265:DNA Helicases / Q000235:genetics / Q000302:isolation & purification* / Q000378:metabolism; D019084:Fluorescent Antibody Technique, Indirect; D006680:HLA Antigens / Q000235:genetics*; D006367:HeLa Cells; D006801:Humans; D008285:Major Histocompatibility Complex / Q000235:genetics*; D008970:Molecular Weight; D009687:Nuclear Proteins / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism; D011132:Polyribosomes / Q000737:chemistry; D011485:Protein Binding; D014176:Protein Biosynthesis; D012333:RNA, Messenger / Q000032:analysis / Q000378:metabolism; D011993:Recombinant Fusion Proteins / Q000302:isolation & purification / Q000378:metabolism; D012269:Ribosomal Proteins / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism; D012270:Ribosomes / Q000737:chemistry*; D014018:Tissue Distribution
D006680:HLA Antigens; D009687:Nuclear Proteins; D012333:RNA, Messenger; D011993:Recombinant Fusion Proteins; D012269:Ribosomal Proteins; D004265:DNA Helicases; C114718:SKIV2L protein, human