CDNA sequence and tissue distribution of the mRNA for bovine and murine p11, the S100-related light chain of the protein-tyrosine kinase substrate p36 (calpactin I).

C J Saris,T Kristensen,P D'Eustachio,L J Hicks,D J Noonan,T Hunter,B F Tack

doi:10.1016/s0021-9258(18)61015-4

Abstract

We have isolated and sequenced cDNA clones of bovine and murine p11 mRNAs. The nonpolyadenylated mRNAs are predicted to be 614 and 600 nucleotides, respectively. The p11 mRNAs both contain a 291 nucleotide open reading frame, preceded by a 5'-untranslated region of 73 nucleotides in bovine p11 mRNA and of 68 nucleotides in murine p11 mRNA. The deduced bovine p11 amino acid sequence is identical to the previously published partial bovine and complete porcine p11 protein sequence except for an additional COOH-terminal lysine residue. The bovine and murine p11 proteins are 92% homologous, whereas at the nucleotide level the conservation is 89% in the coding region and 75% in the 3'-untranslated region. Southern analysis of murine genomic DNA detected a single p11 gene, less than 10 kilobase pairs in size, containing as many as three introns. The p11 gene has been assigned to mouse chromosome 3 by analysis of interspecific hybrid cell panels and recombinant inbred mouse strains. The p11 gene is closely linked to the Xmmv-65 endogenous leukemia virus env gene and the guanylate binding protein-1 gene. Northern analyses of RNAs from mouse tissues and cell lines indicated that p11 mRNA levels vary widely. They are very low in liver, heart, and testes, moderate in brain, spleen, and thymus, and high in kidney, intestine, and lung. Analysis of the same RNA samples for p36 mRNA levels showed that expression of p11 and p36 mRNAs is not always coordinated. Brain and the mouse embryonal carcinoma cell line F9 contain moderate to high levels of p11 mRNA with very low levels of p36 mRNA. Sequence homology between p11 and the S100 proteins, and the serum-induced 2A9 gene product, as well as possible functions of p11 are discussed.

Highlights

MRNAs are predicted to be 614 and 600 nucleotides, kinase C, is a protein of 38 kDa, variously referred to asp34, respectively
Brain and the mouse embryonal carcinoma cell line F9 contain moderate to high levels of p l l mRNA with very low levels of p36 mRNA
The location of the major proteolytic cleavage site reflects the overall structure of p36 asit emerges from the sequence, which indicates that the33-kDa core is composed of four repeating elements of approximately 70 amino acids each, preceded by 20 amino acids which donot form part of the repeat motif [8, 10]

Summary

RESULTS

3‘-untranslated regions of 250 and 241 nucleotides, this cDNACloning and Sequencing-An Okayama-Berg [36] makes the nonpolyadenylated p l l bovine and murine mRNAs cDNA library of RNA from bovine intestinal epithelial cells 614 and 600 nucleotides, respectively. Since the ATG codon was constructed, plated induplicate on nitrocellulose, and the at positions 69-71 in the murine cDNA sequence is the first filters were hybridized to either oligonucleotide mix A This 8-14) or to mix B (covering amino acids 34-39) as described location is consistent with the fact that themurine and bovine under “Experimental Procedures.”. The abbreviationsused are: MDBK, Madin-Darbybovine kidney cells; kb, kilobase pair(s); CaBP, Ca2+-bindingprotein; AMV, avian myeloblastosis virus (Miniprint); SDS, sodium dodecyl sulfate (Miniprint)

IIIIIII II

IIIII I I l l IIIIIIIII I I Ill I I IIIIIIIII Ill1 I I I I IIIII IIIII

IIIIIIII IIII IIIIIIIII I l l IIIIIIIIIIIIIII

DISCUSSION

I11 I I I I