Subtraction cloning of H-rev107, a gene specifically expressed in H-ras resistant fibroblasts.
We have isolated by subtractive hybridization a novel gene, called H-rev107, which is specifically expressed in a phenotypic revertant of H-ras transformed 208F rat fibroblasts. Apart from oncogene revertants, strong expression of H-rev107 was found in REF52 and EK-3 cells, two fibroblast lines resistant to transformation by activated H-ras oncogenes. In contrast, transformation-sensitive fibroblasts like 208F or NIH3T3 cells expressed only very little H-rev107 RNA. In H-ras or v-src transformed fibroblasts, H-rev107 RNA was undetectable. Introduction of the adenovirus E1A nuclear oncogene into ras-resistant REF52 cells abolished their transformation resistance and repressed the H-rev107 gene. H-rev107 encodes a protein with a molecular weight of 18 kDa without any structural similarity to known proteins. p18H-rev107 exists in two forms which can be distinguished by their electrophoretic mobility; one is localized predominantly in cell membranes, the other in the cytoplasm. In confluent contact-inhibited 208F cells, p18H-rev107 accumulated in cell membranes, while growth arrest induced by serum starvation did not induce H-rev107. In REF52, cell density had no influence on the expression or localization of p18H-rev107. Repression of the H-rev107 gene may be closely associated with the loss of density-dependent growth inhibition and with the expression of the neoplastic phenotype.
Hajnal
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Department of Pathology, University of Zürich, Switzerland.
Klemenz
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Schäfer
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D017500:Adenovirus E1A Proteins / Q000502:physiology; D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D015152:Blotting, Northern; D015153:Blotting, Western; D002460:Cell Line; D002462:Cell Membrane / Q000737:chemistry; D002471:Cell Transformation, Neoplastic / Q000235:genetics* / Q000473:pathology; D003001:Cloning, Molecular; D003593:Cytoplasm / Q000737:chemistry; D004247:DNA / Q000032:analysis / Q000235:genetics; D015536:Down-Regulation / Q000502:physiology; D005347:Fibroblasts / Q000737:chemistry / Q000378:metabolism / Q000473:pathology*; D011905:Genes, ras / Q000235:genetics / Q000502:physiology*; D008969:Molecular Sequence Data; D008970:Molecular Weight; D010641:Phenotype; D054512:Phospholipases A2, Calcium-Independent; D011506:Proteins / Q000032:analysis / Q000235:genetics* / Q000378:metabolism; D012333:RNA, Messenger / Q000032:analysis / Q000235:genetics; D051381:Rats; D025521:Tumor Suppressor Proteins
D017500:Adenovirus E1A Proteins; D011506:Proteins; D012333:RNA, Messenger; D025521:Tumor Suppressor Proteins; D004247:DNA; D054512:Phospholipases A2, Calcium-Independent; C085466:Plaat3 protein, rat