Isolation and characterization of a novel PDGF-induced human gene.
Using a differential display RT-PCR strategy to identify novel growth-factor-induced transcripts, we cloned and characterized the human homolog of yeast NOP5/NOP58, whose gene product has been implicated in the execution of early pre-rRNA processing steps. Human NOP5 cDNA was isolated from an M426 fibroblast cDNA library. Determination of the cDNA nucleotide sequence revealed an open reading frame of 1587 nucleotides encoding a predicted gene product of 529 amino acids and mass of 59554Da. The yeast and human NOP5 gene products were found to share 63% homology and 46% identity. NOP5 mRNA was induced within 2h of platelet-derived growth factor (PDGF) treatment of human M426 fibroblasts. Pretreatment with cycloheximide enhanced, while actinomycin blocked induction of the NOP5 transcript. In vitro translational analysis of the cDNA revealed a 60kDa species, consistent with the predicted molecular weight of the gene product. Ubiquitous, but differential NOP5 mRNA expression was revealed after Northern blot analysis of total RNA from several human tissues. Moreover, NOP5 mRNA expression was also demonstrated in cell lines of fibroblast, epithelial, and myeloid origin. A highly charged carboxy terminal domain and consensus phosphorylation sites were identified. The presence of potential regulatory elements, together with growth factor induction and widespread expression is consistent with the hypothesis that the NOP5 gene product may play a role in fundamental cellular growth processes.
Laboratory of Cellular and Molecular Biology, National Cancer Institute, Building 37, Room 1E24, Bethesda, MD 20892, USA.
S A
SA
Nelson
K E
KE
Santora
W J
WJ
LaRochelle
D000595:Amino Acid Sequence; D001483:Base Sequence; D015152:Blotting, Northern; D002460:Cell Line; D018076:DNA, Complementary / Q000737:chemistry / Q000235:genetics / Q000302:isolation & purification*; D005260:Female; D005347:Fibroblasts / Q000166:cytology / Q000187:drug effects / Q000378:metabolism; D005786:Gene Expression Regulation / Q000187:drug effects; D006801:Humans; D020014:K562 Cells; D008969:Molecular Sequence Data; D009687:Nuclear Proteins; D010982:Platelet-Derived Growth Factor / Q000494:pharmacology*; D014176:Protein Biosynthesis; D012333:RNA, Messenger / Q000235:genetics / Q000378:metabolism; D020718:Ribonucleoproteins, Small Nucleolar / Q000235:genetics*; D029701:Saccharomyces cerevisiae Proteins; D017422:Sequence Analysis, DNA; D014018:Tissue Distribution; D014407:Tumor Cells, Cultured
D018076:DNA, Complementary; C438199:NOP58 protein, S cerevisiae; C499062:NOP58 protein, human; D009687:Nuclear Proteins; D010982:Platelet-Derived Growth Factor; D012333:RNA, Messenger; D020718:Ribonucleoproteins, Small Nucleolar; D029701:Saccharomyces cerevisiae Proteins