Isolation and sequence of a cDNA clone for human calcineurin B, the Ca2+-binding subunit of the Ca2+/calmodulin-stimulated protein phosphatase.

We have identified and cloned human cDNA for the Ca2+-binding subunit of calcineurin, the brain isozyme of the Ca2+/calmodulin-stimulated protein phosphatase. The 2.5-kb cDNA has an open reading frame of 510 bp, a leader sequence of at least 500 bp, and a 1,277-bp 3'-noncoding sequence. The deduced sequence of the human protein differs from bovine brain calcineurin B by an additional valine at the carboxyl terminus and substitution of Met-11 and Ser-153 by cysteine. A partial clone of the mouse protein corresponding to amino acids 75-150 was also isolated. This portion of the human and mouse protein sequence is identical, with the DNA sequences showing 94% identity. The respective mRNAs in human and mouse are also of similar size. As was observed with protein levels, mRNA abundance in brain is 20-60 times that found in other tissues with the exception of HeLa cells which, like brain, contain abundant calcineurin B mRNA.

[1]  G. Fink,et al.  A suppressor of a HIS4 transcriptional defect encodes a protein with homology to the catalytic subunit of protein phosphatases , 1989, Cell.

[2]  R. Evans,et al.  Phosphorylation of vimentin in mitotically selected cells. In vitro cyclic AMP-independent kinase and calcium-stimulated phosphatase activities , 1989, The Journal of cell biology.

[3]  T. Sugimura,et al.  Molecular cloning of rat phosphoprotein phosphatase 2Aβ cDNA and increased expressions of phosphatase 2Aα and 2Aβ in rat liver tumors , 1988 .

[4]  M. Kozak.  An analysis of 5'-noncoding sequences from 699 vertebrate messenger RNAs. , 1987, Nucleic acids research.

[5]  C. Richardson,et al.  DNA sequence analysis with a modified bacteriophage T7 DNA polymerase. , 1987, Proceedings of the National Academy of Sciences of the United States of America.

[6]  J. Sikela,et al.  Screening an expression library with a ligand probe: isolation and sequence of a cDNA corresponding to a brain calmodulin-binding protein. , 1987, Proceedings of the National Academy of Sciences of the United States of America.

[7]  D. Towler,et al.  Amino-terminal processing of proteins by N-myristoylation. Substrate specificity of N-myristoyl transferase. , 1987, The Journal of biological chemistry.

[8]  J. Kamholz,et al.  Identification of three forms of human myelin basic protein by cDNA cloning. , 1986, Proceedings of the National Academy of Sciences of the United States of America.

[9]  F. Seela,et al.  Improvement of the dideoxy chain termination method of DNA sequencing by use of deoxy-7-deazaguanosine triphosphate in place of dGTP. , 1986, Nucleic acids research.

[10]  M. Hattori,et al.  Dideoxy sequencing method using denatured plasmid templates. , 1986, Analytical biochemistry.

[11]  W. Y. Cheung,et al.  Bovine brain calmodulin-dependent protein phosphatase. Regulation of subunit A activity by calmodulin and subunit B. , 1985, The Journal of biological chemistry.

[12]  S. Henikoff.  Unidirectional digestion with exonuclease III creates targeted breakpoints for DNA sequencing. , 1984, Gene.

[13]  P. Cohen,et al.  The structure of the B subunit of calcineurin. , 1984, European journal of biochemistry.

[14]  R. W. Davis,et al.  Yeast RNA polymerase II genes: isolation with antibody probes. , 1983, Science.

[15]  G. Hong,et al.  Buffer gradient gels and 35S label as an aid to rapid DNA sequence determination. , 1983, Proceedings of the National Academy of Sciences of the United States of America.

[16]  P. Cohen,et al.  The protein phosphatases involved in cellular regulation. 6. Measurement of type-1 and type-2 protein phosphatases in extracts of mammalian tissues; an assessment of their physiological roles. , 1983, European journal of biochemistry.

[17]  P. Cohen,et al.  The protein phosphatases involved in cellular regulation. 5. Purification and properties of a Ca2+/calmodulin-dependent protein phosphatase (2B) from rabbit skeletal muscle. , 1983, European journal of biochemistry.

[18]  R. Cortese,et al.  pEMBL: a new family of single stranded plasmids. , 1983, Nucleic acids research.

[19]  C. Klee,et al.  Identification of the NH2‐terminal blocking group of calcineurin B as myristic acid , 1982, FEBS letters.

[20]  P. Cohen,et al.  Discovery of A Ca2+‐and calmodulin‐dependent protein phosphatase , 1982, FEBS letters.

[21]  D. Holmes,et al.  A rapid boiling method for the preparation of bacterial plasmids. , 1981, Analytical biochemistry.

[22]  R W Wallace,et al.  High levels of a heat-labile calmodulin-binding protein (CaM-BP80) in bovine neostriatum. , 1980, Biochemistry.

[23]  C. Klee,et al.  Calcineurin: a calcium- and calmodulin-binding protein of the nervous system. , 1979, Proceedings of the National Academy of Sciences of the United States of America.

[24]  S. L. Berger,et al.  Inhibition of intractable nucleases with ribonucleoside--vanadyl complexes: isolation of messenger ribonucleic acid from resting lymphocytes. , 1979, Biochemistry.

[25]  F. Sanger,et al.  DNA sequencing with chain-terminating inhibitors. , 1977, Proceedings of the National Academy of Sciences of the United States of America.

[26]  W. D. Benton,et al.  Screening lambdagt recombinant clones by hybridization to single plaques in situ. , 1977, Science.