Two novel variants of the v-src oncogene isolated from low and high metastatic RSV-transformed hamster cells.

Tatosyan A, Yatsula B, Shtutman M, Moinova E, Kaverina I, Musatkina E, Leskov K, Mizenina O, Zueva E, Calothy G, Dezélée P
Virology. 1996 216 (2): 347-56

PMID: 8607264 · DOI:10.1006/viro.1996.0070

Four different transformed cell lines were isolated as a result of independent infection of primary hamster fibroblasts by Rous sarcoma virus (RSV SR-D stocks). These lines differ by the level of their spontaneous metastatic activity: HET-SR-1, HET-SR-8, and HET-SR-10 cell lines induced 70-200 metastatic nodules in the lung and/or lymph nodes of inoculated animals (high metastatic lines, HM). Metastatic activity was not identified after injection of HET-SR cells (low metastatic line, LM). All cell lines contained one copy of integrated and expressed intact RSV provirus. The difference in the amount of v-src protein in cell lines was not correlated with their metastatic potential in vivo. Complete v-srcHM and v-srcLM genes were cloned from corresponding gene libraries and sequenced. In the unique region of both v-src isoforms a GC-rich insert of 60 nucleotides (20 a.a.) was found. The presence of this insert explains the unusual apparent molecular weight of protein encoded by v-srcHM and vsrcLM: 62 kDA. Both genes had 10 identical amino acid changes when compared to the known RSV SR-D v-src sequence. v-srcHM and v-srcLM differ by several amino acid changes. Most of them are localized in the unique domain and the extreme carboxy-terminal region of the of the oncoprotein. Both v-src variants and chimeric v-src with mutually substituted parts were subcloned in a retroviral vector and introduced into avian neuroretina cells. Significant differences in the morphology of transformed neuroretinal cells were associated with the mutations in the carboxy-terminal region of the v-src oncogene. Low metastatic HET-SR cells transfected with v-srcHM and the chimeric gene v-src-LH remarkably increased their metastatic potential. In contrast, this effect was not observed when the same cells were transfected with v-srcLM and the chimeric v-srcHL gene. Specific changes in the distribution of fibronectin matrix typical for high metastatic cells were found in the lines transfected with v-srcHM.

MeSH Terms (22)

Amino Acid Sequence Animals Avian Sarcoma Viruses Base Sequence Cell Line, Transformed Cell Transformation, Neoplastic Cell Transformation, Viral Chickens Cloning, Molecular Cricetinae DNA, Viral Fibronectins Genes, src Genes, Viral Mesocricetus Molecular Sequence Data Mutation Neoplasm Metastasis Neoplasms, Experimental Plasmids Recombinant Fusion Proteins Retina

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