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PMID: 2122236 Published · ppublish English Comparative Study Journal Article

Molecular cloning and expression of a G25K cDNA, the human homolog of the yeast cell cycle gene CDC42.

Molecular and cellular biology ·Vol. 10 ·No. 11 ·1990-11-00 ·Pages 5977-82

Munemitsu S, Innis MA, Clark R, McCormick F, Ullrich A, Polakis P

Abstract

G25K is a low-molecular-mass GTP-binding protein with a broad distribution in mammalian tissues. A cDNA clone was isolated by using oligonucleotides corresponding to the partial amino acid sequence of purified human G25K. The cDNA encodes an 191-amino-acid polypeptide containing GTP-binding consensus sequences and a putative farnesylation site at the C terminus. The sequence exhibits 50 and 70% identities to the mammalian rho and rac proteins, respectively, and an 80% identity to the Saccharomyces cerevisiae CDC42 gene product. Insect Sf9 cells infected with recombinant baculovirus vectors expressing the G25K cDNA produced a 25-kDa protein that bound GTP and was recognized by antibodies specifically reactive to G25K. G25K appears to be the human homolog of the CDC42 gene product, since expression of the G25K cDNA in S. cerevisiae suppressed both cdc42-1 and cdc24-4 temperature-sensitive lethal mutations.

MeSH Terms
Amino Acid Sequence Base Sequence Cell Cycle Cloning, Molecular/methods Fungal Proteins/genetics GTP-Binding Proteins/genetics Genes, Fungal Humans Molecular Sequence Data Oligonucleotide Probes Plasmids Polymerase Chain Reaction Saccharomyces cerevisiae/cytology,genetics Sequence Homology, Nucleic Acid Temperature cdc42 GTP-Binding Protein cdc42 GTP-Binding Protein, Saccharomyces cerevisiae
Chemicals
Fungal Proteins Oligonucleotide Probes GTP-Binding Proteins cdc42 GTP-Binding Protein cdc42 GTP-Binding Protein, Saccharomyces cerevisiae
Authors & Affiliations
6 authors, click to expand affiliations / ORCID
Munemitsu S
Department of Molecular Biology, Cetus Corporation, Emeryville, California 94608.
Innis M A
Clark R
McCormick F
Ullrich A
Polakis P
References (44)
44 references, click to expand
  1. Cleavage of structural proteins during the assembly of the head of bacteriophage T4.
    Nature. 1970 Aug 15;227(5259):680-5 PMID: 5432063
  2. ras p21 protein promotes survival and fiber outgrowth of cultured embryonic neurons.
    Neuron. 1989 Jan;2(1):1087-96 PMID: 2696501
  3. DNA sequencing with chain-terminating inhibitors.
    Proc Natl Acad Sci U S A. 1977 Dec;74(12):5463-7 PMID: 271968
  4. A system for shotgun DNA sequencing.
    Nucleic Acids Res. 1981 Jan 24;9(2):309-21 PMID: 6259625
  5. The guanine nucleotide activating site of the regulatory component of adenylate cyclase. Identification by ligand binding.
    J Biol Chem. 1982 Oct 10;257(19):11416-23 PMID: 6288684
  6. Production of human beta interferon in insect cells infected with a baculovirus expression vector.
    Mol Cell Biol. 1983 Dec;3(12):2156-65 PMID: 6318086
  7. A general method for polyethylene-glycol-induced genetic transformation of bacteria and yeast.
    Gene. 1983 Nov;25(2-3):333-41 PMID: 6363214
  8. Relationship of actin and tubulin distribution to bud growth in wild-type and morphogenetic-mutant Saccharomyces cerevisiae.
    J Cell Biol. 1984 Mar;98(3):934-45 PMID: 6365931
  9. Isolation of the human insulin-like growth factor I gene using a single synthetic DNA probe.
    EMBO J. 1984 Feb;3(2):361-4 PMID: 6232133
  10. Synthetic oligonucleotide probes deduced from amino acid sequence data. Theoretical and practical considerations.
    J Mol Biol. 1985 May 5;183(1):1-12 PMID: 4009718
  11. Inhibition of growth factor-induced differentiation of PC12 cells by microinjection of antibody to ras p21.
    Nature. 1986 Feb 20-26;319(6055):680-2 PMID: 3005866
  12. Purification of the major GTP-binding proteins from human placental membranes.
    J Biol Chem. 1986 May 25;261(15):7052-9 PMID: 3084494
  13. The oncogenic activation of human p21ras by a novel mechanism.
    Science. 1986 Aug 8;233(4764):649-52 PMID: 3487832
  14. Ras p21 as a potential mediator of insulin action in Xenopus oocytes.
    Science. 1987 May 15;236(4803):840-3 PMID: 3554510
  15. Guanine nucleotide activation of, and competition between, RAS proteins from Saccharomyces cerevisiae.
    Mol Cell Biol. 1987 Jun;7(6):2128-33 PMID: 3299060
  16. Nucleotide sequence of the CLS4 (CDC24) gene of Saccharomyces cerevisiae.
    Gene. 1987;54(1):125-32 PMID: 3301539
  17. A cytoplasmic protein stimulates normal N-ras p21 GTPase, but does not affect oncogenic mutants.
    Science. 1987 Oct 23;238(4826):542-5 PMID: 2821624
  18. Identification and purification from bovine brain of a guanine-nucleotide-binding protein distinct from Gs, Gi and Go.
    Biochem J. 1987 Sep 1;246(2):431-9 PMID: 3120696
  19. Primer-directed enzymatic amplification of DNA with a thermostable DNA polymerase.
    Science. 1988 Jan 29;239(4839):487-91 PMID: 2448875
  20. Induction of neurite formation in PC12 cells by microinjection of proto-oncogenic Ha-ras protein preincubated with guanosine-5'-O-(3-thiotriphosphate).
    Mol Cell Biol. 1987 Dec;7(12):4553-6 PMID: 3325827
  21. The yeast GTP-binding YPT1 protein and a mammalian counterpart are associated with the secretion machinery.
    Cell. 1988 Mar 25;52(6):915-24 PMID: 3127057
  22. A GTP-binding protein required for secretion rapidly associates with secretory vesicles and the plasma membrane in yeast.
    Cell. 1988 Jun 3;53(5):753-68 PMID: 3131018
  23. A ras-related gene with transformation suppressor activity.
    Cell. 1989 Jan 13;56(1):77-84 PMID: 2642744
  24. ADP-ribosylation of the rho/rac proteins induces growth inhibition, neurite outgrowth and acetylcholine esterase in cultured PC-12 cells.
    Biochem Biophys Res Commun. 1990 Feb 28;167(1):265-72 PMID: 2106882
  25. The identification and characterization of an epidermal growth factor-stimulated phosphorylation of a specific low molecular weight GTP-binding protein in a reconstituted phospholipid vesicle system.
    J Biol Chem. 1990 Apr 15;265(11):5990-6001 PMID: 2156836
  26. Biochemical characterization of baculovirus-expressed rap1A/Krev-1 and its regulation by GTPase-activating proteins.
    Mol Cell Biol. 1990 Jun;10(6):2901-8 PMID: 2160589
  27. CDC42 and CDC43, two additional genes involved in budding and the establishment of cell polarity in the yeast Saccharomyces cerevisiae.
    J Cell Biol. 1990 Jul;111(1):131-42 PMID: 2195038
  28. Molecular characterization of CDC42, a Saccharomyces cerevisiae gene involved in the development of cell polarity.
    J Cell Biol. 1990 Jul;111(1):143-52 PMID: 2164028
  29. A guide to low molecular weight GTPases.
    Cell Growth Differ. 1990 May;1(5):251-8 PMID: 2150754
  30. High level expression of nonfused foreign genes with Autographa californica nuclear polyhedrosis virus expression vectors.
    Virology. 1989 May;170(1):31-9 PMID: 2497580
  31. Mutagenesis of bacterial elongation factor Tu at lysine 136. A conserved amino acid in GTP regulatory proteins.
    J Biol Chem. 1989 May 15;264(14):8304-9 PMID: 2498311
  32. Asparagine residue in the rho gene product is the modification site for botulinum ADP-ribosyltransferase.
    J Biol Chem. 1989 May 25;264(15):8602-5 PMID: 2498316
  33. All ras proteins are polyisoprenylated but only some are palmitoylated.
    Cell. 1989 Jun 30;57(7):1167-77 PMID: 2661017
  34. The mammalian G protein rhoC is ADP-ribosylated by Clostridium botulinum exoenzyme C3 and affects actin microfilaments in Vero cells.
    EMBO J. 1989 Apr;8(4):1087-92 PMID: 2501082
  35. Bombesin stimulation of inositol 1,4,5-trisphosphate generation and intracellular calcium release is amplified in a cell line overexpressing the N-ras proto-oncogene.
    Biochem J. 1989 Jun 15;260(3):813-9 PMID: 2669738
  36. rac, a novel ras-related family of proteins that are botulinum toxin substrates.
    J Biol Chem. 1989 Oct 5;264(28):16378-82 PMID: 2674130
  37. Structure of the guanine-nucleotide-binding domain of the Ha-ras oncogene product p21 in the triphosphate conformation.
    Nature. 1989 Sep 21;341(6239):209-14 PMID: 2476675
  38. Glycosylation of heterologous proteins in Saccharomyces cerevisiae.
    Biotechnology. 1989;13:233-46 PMID: 2679927
  39. Low molecular weight GTP-binding proteins are associated with neuronal organelles involved in rapid axonal transport and exocytosis.
    J Biol Chem. 1989 Nov 5;264(31):18363-7 PMID: 2478544
  40. Genetic analysis of mammalian GAP expressed in yeast.
    Cell. 1989 Nov 17;59(4):681-6 PMID: 2684416
  41. Multicopy suppression of the cdc24 budding defect in yeast by CDC42 and three newly identified genes including the ras-related gene RSR1.
    Proc Natl Acad Sci U S A. 1989 Dec;86(24):9976-80 PMID: 2690082
  42. ADP-ribosylation factor is functionally and physically associated with the Golgi complex.
    Proc Natl Acad Sci U S A. 1990 Feb;87(3):1238-42 PMID: 2105501
  43. Molecular switch for signal transduction: structural differences between active and inactive forms of protooncogenic ras proteins.
    Science. 1990 Feb 23;247(4945):939-45 PMID: 2406906
  44. A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding.
    Anal Biochem. 1976 May 7;72:248-54 PMID: 942051
Article Info
Journal
Molecular and cellular biology
Abbr.
Mol Cell Biol
ISSN
0270-7306
DOI
10.1128/mcb.10.11.5977-5982.1990
Published
1990-11-00
Pages
5977-82
Language
English
Region
United States
NLM ID
8109087
PMCID
PMC361395
Subset
IM
Databases
GENBANK
M35543
External Links
PubMed source DOI: 10.1128/mcb.10.11.5977-5982.1990 PMC Full Text View journal details
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