Isolation of cDNAs encoding a substrate for protein kinase C: Nucleotide sequence and chromosomal mapping of the gene for a human 80K protein

Kosuke Sakai, Masamichi Hirai, Shinsei Minoshima, Jun Kudo, Ryuichi Fukuyama, Nobuyoshi Shimizu

Research output: Contribution to journalArticle

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Abstract

An acidic phosphoprotein of Mr 80,000, the 80K protein, is a substrate for protein kinase C in fibroblasts and epidermal carcinoma cells. We purified the 80K protein from human squamous carcinoma Ca9-22 cells and fractionated it into two distinct molecular species, designated the 80K-L and 80K-H proteins. The amino acid sequences of the NH2-terminal region and cyanogen bromide-cleaved fragments of the 80K-H protein were determined and a corresponding oligonucleotide sequence was synthesized. Using this as a probe, two cDNA clones, λ80H-1 and λ80H-2, were selected from a λgt10 cDNA library from human A431 cells. The nucleotide sequence has an open reading frame of 1581 nucleotides encoding a protein of 527 amino acids. The deduced amino acid sequence revealed an extremely Glu-rich region. RNA blot analysis with the λ80H-1 cDNA clone detected two polyadenylated transcripts of 2.3 and 3.5 kb in Ca9-22 cells. Spot blot hybridization using flow-sorted human chromosomes provided evidence that the gene (G19P1) encoding 80K-H protein maps to human chromosome 19.

Original languageEnglish
Pages (from-to)309-315
Number of pages7
JournalGenomics
Volume5
Issue number2
DOIs
Publication statusPublished - 1989

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Chromosome Mapping
Protein Kinase C
Complementary DNA
Human Chromosomes
Proteins
Amino Acid Sequence
Clone Cells
Chromosomes, Human, Pair 19
Cyanogen Bromide
Phosphoproteins
Gene Library
Oligonucleotides
Open Reading Frames
Squamous Cell Carcinoma
Nucleotides
Fibroblasts
human PRKCSH protein
RNA
Carcinoma
Amino Acids

ASJC Scopus subject areas

  • Genetics

Cite this

Isolation of cDNAs encoding a substrate for protein kinase C : Nucleotide sequence and chromosomal mapping of the gene for a human 80K protein. / Sakai, Kosuke; Hirai, Masamichi; Minoshima, Shinsei; Kudo, Jun; Fukuyama, Ryuichi; Shimizu, Nobuyoshi.

In: Genomics, Vol. 5, No. 2, 1989, p. 309-315.

Research output: Contribution to journalArticle

Sakai, Kosuke ; Hirai, Masamichi ; Minoshima, Shinsei ; Kudo, Jun ; Fukuyama, Ryuichi ; Shimizu, Nobuyoshi. / Isolation of cDNAs encoding a substrate for protein kinase C : Nucleotide sequence and chromosomal mapping of the gene for a human 80K protein. In: Genomics. 1989 ; Vol. 5, No. 2. pp. 309-315.
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