Alternatively spliced murine lyn mRNAs encode distinct proteins.
Two lyn proteins of 56 and 53 kDa have been observed in immunoprecipitates from a variety of murine and human cell lines and tissues. We report the cloning and nucleotide sequence of two distinct murine lyn cDNAs isolated from an FDC-P1 cDNA library. One of the cDNAs, designated lyn11, encodes a protein of 56 kDa which shares 96% similarity with human lyn. The other cDNA, designated lyn12, encodes a protein of 53 kDa. The proteins differ in the presence or absence of a 21-amino-acid sequence located 24 amino acids C terminal of the translational initiation codon. Using RNase protection analysis, we have identified mRNAs corresponding to both cDNAs in murine cell lines and tissues. Sequence analysis of murine genomic clones suggests that the distinct mRNAs are alternatively spliced transcripts derived from a single gene. Expression of both cDNAs in COS cells leads to the production of lyn proteins with the same molecular weight as the two forms of lyn proteins immunoprecipitated from extracts of FDC-P1 cells and mouse spleen. Subcellular fractionation studies and Western immunoblotting analysis suggest that both isoforms of lyn are membrane associated. The association of both lyn isoforms with the membrane fraction supports the notion that lyn, like other src-related kinases, may interact with the intracellular domain of cell surface receptors.
Stanley
E
E
Melbourne Tumour Biology Branch, Ludwig Institute for Cancer Research, Parkville, Victoria, Australia.
Ralph
S
S
McEwen
S
S
Boulet
I
I
Holtzman
D A
DA
Lock
P
P
Dunn
A R
AR
D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D015152:Blotting, Northern; D015139:Blotting, Southern; D002460:Cell Line; D003001:Cloning, Molecular; D016391:Genes, src; D051379:Mice; D008969:Molecular Sequence Data; D008970:Molecular Weight; D015345:Oligonucleotide Probes; D009856:Oncogene Proteins, Viral / Q000235:genetics; D011061:Poly A / Q000235:genetics / Q000302:isolation & purification; D011505:Protein-Tyrosine Kinases / Q000235:genetics*; D012313:RNA / Q000235:genetics / Q000302:isolation & purification; D012326:RNA Splicing; D012333:RNA, Messenger / Q000235:genetics*; D014162:Transfection; D019061:src-Family Kinases
D015345:Oligonucleotide Probes; D009856:Oncogene Proteins, Viral; D012333:RNA, Messenger; D011061:Poly A; D012313:RNA; D011505:Protein-Tyrosine Kinases; C060902:lyn protein-tyrosine kinase; D019061:src-Family Kinases