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PMID: 2026147 Published · ppublish English Comparative Study Journal Article Research Support, Non-U.S. Gov't Research Support, U.S. Gov't, P.H.S.

Multiple Ca2+/calmodulin-dependent protein kinase genes in a unicellular eukaryote.

The EMBO journal ·Vol. 10 ·No. 6 ·1991-06-00 ·Pages 1511-22

Pausch MH, Kaim D, Kunisawa R, Admon A, Thorner J

Abstract

We purified a Ca2+/calmodulin (CaM)-dependent protein kinase (CaM kinase) from the yeast Saccharomyces cerevisiae with properties similar to mammalian type II CaM kinases. Degenerate oligonucleotides designed on the basis of the amino acid sequence of tryptic peptides from the 55 kd subunit of the yeast CaM kinase were used to isolate its gene from a set of lambda gt11-yeast genomic DNA phage clones initially selected by the ability to bind 125I-labelled yeast CaM. The cloned gene (CMK1) encodes an open reading frame that is homologous to the sequences of vertebrate type II CaM kinases. Several criteria demonstrated that the CMK1 gene product is the 55 kd polypeptide. Neither over-production (11-fold) nor complete elimination of the CMK1 gene product had any detectably deleterious effect on yeast cell growth. Extracts from cmk1 delta cells, which lacked detectable p55 using an antiserum raised against a Staphylococcus aureus protein A-CMK1 fusion protein, possessed significant residual Ca2+/CAM-dependent protein kinase activity. Using the CMK1 gene as a probe at low stringency, a second gene (CMK2) encoding another CaM-dependent protein kinase with striking sequence similarity to CMK1 was cloned. Deletion of CMK2, or both CMK1 and CMK2, was not lethal, although loss of CMK2 caused a slow rate of spore germination.

Related Genes
CMK1 CMK2
MeSH Terms
Amino Acid Sequence Base Sequence Binding Sites Calcium-Calmodulin-Dependent Protein Kinases/genetics Calmodulin-Binding Proteins/genetics Chromosome Mapping Cloning, Molecular Fungal Proteins/genetics Genes, Fungal Molecular Sequence Data Oligonucleotides/chemistry Oligopeptides/chemistry Protein Kinases/genetics Restriction Mapping Saccharomyces cerevisiae/genetics Saccharomyces cerevisiae Proteins/genetics
Chemicals
Calmodulin-Binding Proteins Fungal Proteins Oligonucleotides Oligopeptides Saccharomyces cerevisiae Proteins Protein Kinases CMK2 protein, S cerevisiae Calcium-Calmodulin-Dependent Protein Kinases
Authors & Affiliations
5 authors, click to expand affiliations / ORCID
Pausch M H
Division of Biochemistry and Molecular Biology, University of California, Berkeley 94720.
Kaim D
Kunisawa R
Admon A
Thorner J
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Article Info
Journal
The EMBO journal
Abbr.
EMBO J
ISSN
0261-4189
Published
1991-06-00
Pages
1511-22
Language
English
Region
England
NLM ID
8208664
PMCID
PMC452815
Subset
IM
Grants
NCI NIH HHS · CA09041 · United States
NIGMS NIH HHS · GM12730 · United States
NIGMS NIH HHS · GM21841 · United States
Databases
GENBANK
M55400, M55401, M55402, M65224, S60919, S60920, X56961, X57782, X59082, X59871
External Links
PubMed source PMC Full Text View journal details
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