The c-cbl proto-oncogene is preferentially expressed in thymus and testis tissue and encodes a nuclear protein

Cas NS-1 is an acutely transforming murine retrovirus that induces early B-lineage lymphomas and occasional myeloid leukemias. The transforming sequence of this virus, v-cbl, shows no homology to known oncogenes but has some similarities to the yeast transcriptional factor GCN4. In this study we used a v-cbl probe to analyze mRNAs from a wide range of murine and human hemopoietic tumor cell lines and detected an 11-kilobase mRNA in all lineages. In normal mouse tissues the expression of c-cbl was highest in testis and thymus tissues, the predominant species in testis tissue being a 3.5-kilobase mRNA. The v-cbl oncogene was inserted into a bacterial expression vector to produce protein for the immunization of rabbits. Affinity-purified v-cbl antibodies identified abundant levels of p100gag-cbl in Cas NS-1-transformed fibroblasts and lower levels of a 135-kilodalton protein (p135c-cbl) in both normal and transformed cells. Subcellular fractionation showed that p100gag-cbl and p135c-cbl are both located in the nucleus and retained following 420 mM salt extraction. These results indicate that the translational product of a c-cbl is a 135-kilodalton nuclear protein.

[1]  N. Copeland,et al.  Identification of two murine loci homologous to the v-cbl oncogene , 1989, Journal of virology.

[2]  S. Klinken,et al.  v-cbl, an oncogene from a dual-recombinant murine retrovirus that induces early B-lineage lymphomas. , 1989, Proceedings of the National Academy of Sciences of the United States of America.

[3]  D. Smith,et al.  Single-step purification of polypeptides expressed in Escherichia coli as fusions with glutathione S-transferase. , 1988, Gene.

[4]  G. V. Vande Woude,et al.  Proto-oncogene expression in germ cell development. , 1988, Trends in genetics : TIG.

[5]  M. Giorgi,et al.  Expression of cellular protooncogenes in the mouse male germ line: a distinctive 2.4-kilobase pim-1 transcript is expressed in haploid postmeiotic cells. , 1988, Proceedings of the National Academy of Sciences of the United States of America.

[6]  A. Pellicer,et al.  Differential expression of the ras gene family in mice , 1987, Molecular and cellular biology.

[7]  A. E. Sippel,et al.  Subnuclear localization of proteins encoded by the oncogene v-myb and its cellular homolog c-myb , 1986, Molecular and cellular biology.

[8]  J. Yunis,et al.  Prognostic significance of chromosomal abnormalities in acute leukaemias and myelodysplastic syndromes. , 1986, Clinics in haematology.

[9]  G. Evan,et al.  Studies on the interaction of the human c-myc protein with cell nuclei: p62c-myc as a member of a discrete subset of nuclear proteins , 1985, Cell.

[10]  R. Hay.  American Type Culture Collection catalogue of cell lines and hybridomas , 1985 .

[11]  C. Yanisch-Perron,et al.  Improved M13 phage cloning vectors and host strains: nucleotide sequences of the M13mp18 and pUC19 vectors. , 1985, Gene.

[12]  C. Dani,et al.  Post-transcriptional regulation of glyceraldehyde-3-phosphate-dehydrogenase gene expression in rat tissues. , 1984, Nucleic acids research.

[13]  W. Langdon,et al.  Identification of a transforming virus from a lymphoma of a mouse infected with a wild mouse retrovirus. , 1984, Current topics in microbiology and immunology.

[14]  J. Sambrook,et al.  Molecular Cloning: A Laboratory Manual , 2001 .

[15]  R. Roeder,et al.  Accurate transcription initiation by RNA polymerase II in a soluble extract from isolated mammalian nuclei. , 1983, Nucleic acids research.

[16]  I. Verma,et al.  Differential expression of cellular oncogenes during pre- and postnatal development of the mouse , 1982, Nature.

[17]  P. Thomas,et al.  Hybridization of denatured RNA and small DNA fragments transferred to nitrocellulose. , 1980, Proceedings of the National Academy of Sciences of the United States of America.

[18]  D. Kemp,et al.  Expression of the immunoglobulin C mu gene in mouse T and B lymphoid and myeloid cell lines. , 1980, Proceedings of the National Academy of Sciences of the United States of America.

[19]  H. Towbin,et al.  Electrophoretic transfer of proteins from polyacrylamide gels to nitrocellulose sheets: procedure and some applications. , 1979, Proceedings of the National Academy of Sciences of the United States of America.

[20]  I. Miyoshi,et al.  Establishment of an Epstein-Barr virus-determined nuclear antigen-negative human B-cell line from acute lymphoblastic leukemia. , 1977, Journal of the National Cancer Institute.

[21]  U. K. Laemmli,et al.  Cleavage of Structural Proteins during the Assembly of the Head of Bacteriophage T4 , 1970, Nature.