Isolation of chicken cellular DNA sequences with homology to the region of viral oncogenes that encodes the tyrosine kinase domain.

Abstract

A library of chicken genomic DNA was screened for sequences that could hybridize to a cloned DNA fragment containing the transforming gene (v-fps) of Fujinami sarcoma virus. In addition to c-fps, two unique chicken cellular DNA sequences were isolated that hybridized weakly to v-fps. These sequences hybridized with many other viral oncogenes encoding tyrosine kinases, Sequence analysis of the region where homology was detected revealed a region that is highly conserved among the tyrosine kinases both at the nucleotide and amino acid levels. Although we were unable to detect expression of either chicken cellular DNA sequence in a variety of avian tissues, the data suggest the existence of additional members of the tyrosine kinase gene family. Screening genomic libraries for sequences that hybridize weakly to functional regions of other genes may prove useful for the isolation and characterization of additional members of other gene families.Since the discovery that the src gene of Rous sarcoma virus encodes a gene product that possesses a protein kinase activity that is specific for tyrosine (5, 20), a number of other viral transforming genes have also been shown to encode tyrosine-specific protein kinases (2,19). Sequence analysis has revealed a conserved domain in the region of these genes where the enzymatic activity has been mapped (19). These genes include fps of Fujinami sarcoma virus (37), yes of Y73 avian sarcoma virus (24), src of Rous sarcoma virus (36), ros of UR2 avian sarcoma virus (31), erb-B of avian erythroblastosis virus (49), fgr of Gardner-Rasheed feline sarcoma virus (28),fms of Susan McDonough feline sarcoma virus (16), and abl of Abelson murine leukemia virus (34). Other viral oncogenes that do not possess a tyrosine-specific protein kinase activity also share amino acid homology with the tyrosine kinase genes. Among these are mos of Moloney murine sarcoma virus (33, 44), rel of reticuloendotheliosis virus (41), and mil of MH2 (22,42). In addition to the viral oncogenes that have been characterized, other proteins have been isolated that possess tyrosine kinase activity. These include the growth factor receptors for epidermal growth factor (4, 9), for platelet-derived growth factor (11, 17), for insulin-like growth factor-1 (21), and for insulin (10,18,23,43). TPK75 is a tyrosine-specific protein kinase that was isolated from rat liver (48), and NCP94 is a tyrosine-specific protein kinase that was immunoprecipitated from bone marrow and other cells with an antibody raised against a peptide isolated from the tyrosine kinase domain offps (13).The ability to immunologically identify additional tyrosine specific protein kinases (13) suggests that tyrosine kinase genes could be detected by hybridization with DNA probes isolated from the conserved kinase region. Using this approach, we isolated two chicken cellular DNA sequences that likely represent additional members of the tyrosine kinase gene family.Isolation of chicken cellular DNA sequences with homology * Corresponding author. to the tyrosi...

Molecular cloning and expression of chicken C-terminal Src kinase: lack of stable association with c-Src protein.

“…As shown in Fig. 4 B-D, the src probe cross-hybridized with many other genes under lower stringency conditions as expected (28). Under the same conditions, the chicken csk probe revealed one predominant band.…”

Novel tyrosine kinase identified by phosphotyrosine antibody screening of cDNA libraries.

“…Therefore, the isolation and characterization of these genes allow for the identification of proteins which may be important in cellular growth control pathways. While a number of novel tyrosine kinases have been identified by screening cDNA and genomic libraries at low stringency with probes homologous to the catalytic domains of already identified protein tyrosine kinases (9,18,21), we sought to develop new methods for identifying these genes while simultaneously allowing for large-scale production of identified protein kinases and confirmation of the catalytic activity of the cloned genes.…”

Evolution, expression, and chromosomal location of a novel receptor tyrosine kinase gene, eph.

“…As shown in Fig. 2, the pattern of splicing of the eph gene was found to be completely different from that of the other protein tyrosine kinase members, including c-src (1,72), c-fps (28,61), TKRll and TKR16 (21) (not shown), Arg (38), and HER-2 (66). Although it is not shown in Fig.…”