Home LiteratureArticle Details
PMID: 3277185 Published · ppublish English Journal Article

Chimeric proteins define variable and essential regions of Ha-ras-encoded protein.

Proceedings of the National Academy of Sciences of the United States of America ·Vol. 85 ·No. 4 ·1988-02-00 ·Pages 1015-9

Lowe DG, Ricketts M, Levinson AD, Goeddel DV

Abstract

The biological role of amino acid differences between the human 21-kDa Ha-ras protein (p21) and the human 23-kDa R-ras protein (p23) was investigated by engineering mutant Ha-ras p21 molecules containing divergent amino acid sequences from R-ras p23. Variant amino acids from R-ras p23 regions 1-30, 52-57, 67-78, 1-30 and 67-78 together, and 112-124 were substituted for the corresponding Ha-ras p21 amino acid regions 1-4, 26-31, 41-52, 1-4 and 41-52 together, and 86-98, respectively. Rat fibroblasts transfected with genes encoding these position-12 valine-substituted chimeric Ha-ras proteins displayed the same properties of morphological transformation and anchorage-independent growth as Ha-ras T24 oncogene-transformed fibroblasts. However, substitution of variant amino acids from the 80 C-terminal residues (amino acids 138-218) of R-ras p23 for the corresponding p21 amino acids (residues 112-189) inactivated the transforming activity of position-12 valine-substituted p21. The converse substitution of Ha-ras p21 C-terminal residues into R-ras p23 did not result in transformation by position-38 valine-substituted p23. These data are discussed in terms of the structure of ras proteins and the nature of interactions determining the specificity of effector function.

MeSH Terms
Amino Acid Sequence Animals Cell Transformation, Neoplastic Fibroblasts Molecular Sequence Data Protein Conformation Proto-Oncogene Proteins/genetics,physiology Proto-Oncogene Proteins p21(ras) Rats Recombinant Fusion Proteins/genetics,physiology
Chemicals
Proto-Oncogene Proteins Recombinant Fusion Proteins Proto-Oncogene Proteins p21(ras)
Authors & Affiliations
4 authors, click to expand affiliations / ORCID
Lowe D G
Department of Molecular Biology, Genentech, Inc., South San Francisco, CA 94080.
Ricketts M
Levinson A D
Goeddel D V
References (47)
47 references, click to expand
  1. Activation of Ki-ras2 gene in human colon and lung carcinomas by two different point mutations.
    Nature. 1983 Aug 11-17;304(5926):507-13 PMID: 6308467
  2. Heterologous expression and characterization of the human R-ras gene product.
    Mol Cell Biol. 1987 Aug;7(8):2845-56 PMID: 3313005
  3. ras-Related gene sequences identified and isolated from Saccharomyces cerevisiae.
    Nature. 1983 Dec 15-21;306(5944):707-9 PMID: 6318116
  4. Genes in S. cerevisiae encoding proteins with domains homologous to the mammalian ras proteins.
    Cell. 1984 Mar;36(3):607-12 PMID: 6365329
  5. Photoaffinity labeling with GTP of viral p21 ras protein expressed in Escherichia coli.
    J Virol. 1984 May;50(2):343-51 PMID: 6323735
  6. Homologies in the primary structure of GTP-binding proteins: the nucleotide-binding site of EF-Tu and p21.
    EMBO J. 1984 Feb;3(2):339-41 PMID: 6609071
  7. The p21 ras C-terminus is required for transformation and membrane association.
    Nature. 1984 Aug 16-22;310(5978):583-6 PMID: 6087162
  8. Comparative biochemical properties of normal and activated human ras p21 protein.
    Nature. 1984 Aug 23-29;310(5979):644-9 PMID: 6147754
  9. Intrinsic GTPase activity distinguishes normal and oncogenic ras p21 molecules.
    Proc Natl Acad Sci U S A. 1984 Sep;81(18):5704-8 PMID: 6148751
  10. Biological properties of human c-Ha-ras1 genes mutated at codon 12.
    Nature. 1984 Nov 1-7;312(5989):71-5 PMID: 6092966
  11. Harvey murine sarcoma virus p21 ras protein: biological and biochemical significance of the cysteine nearest the carboxy terminus.
    EMBO J. 1984 Nov;3(11):2581-5 PMID: 6096132
  12. Expression of prokaryotic genes for hygromycin B and G418 resistance as dominant-selection markers in mouse L cells.
    Gene. 1984 Oct;30(1-3):147-56 PMID: 6096211
  13. Functional homology of mammalian and yeast RAS genes.
    Cell. 1985 Jan;40(1):19-26 PMID: 2981628
  14. In yeast, RAS proteins are controlling elements of adenylate cyclase.
    Cell. 1985 Jan;40(1):27-36 PMID: 2981630
  15. Requirement for ras proto-oncogene function during serum-stimulated growth of NIH 3T3 cells.
    Nature. 1985 Jan 17-23;313(5999):241-3 PMID: 3918269
  16. Regional homology in GTP-binding proto-oncogene products and elongation factors.
    J Cyclic Nucleotide Protein Phosphor Res. 1983-1984;9(6):435-48 PMID: 6396323
  17. Mammalian and yeast ras gene products: biological function in their heterologous systems.
    Science. 1985 Apr 12;228(4696):179-84 PMID: 3883495
  18. The molecular genetics of cellular oncogenes.
    Annu Rev Genet. 1984;18:553-612 PMID: 6397126
  19. Transient reversion of ras oncogene-induced cell transformation by antibodies specific for amino acid 12 of ras protein.
    Nature. 1985 Apr 18-24;314(6012):639-42 PMID: 2581140
  20. Characterization of functional domains of p21 ras by use of chimeric genes.
    EMBO J. 1985 Feb;4(2):407-12 PMID: 3926484
  21. The ras oncogene product p21 is not a regulatory component of adenylate cyclase.
    Nature. 1985 Sep 5-11;317(6032):71-2 PMID: 3929144
  22. Structure of the GDP domain of EF-Tu and location of the amino acids homologous to ras oncogene proteins.
    Science. 1985 Oct 4;230(4721):32-6 PMID: 3898365
  23. A model for the tertiary structure of p21, the product of the ras oncogene.
    Science. 1985 Oct 4;230(4721):78-82 PMID: 3898366
  24. Microinjection of the ras oncogene protein into PC12 cells induces morphological differentiation.
    Cell. 1985 Oct;42(3):841-8 PMID: 2996779
  25. Sarcoma viruses carrying ras oncogenes induce differentiation-associated properties in a neuronal cell line.
    Nature. 1985 Nov 7-13;318(6041):73-5 PMID: 4058592
  26. The action of oncogenes in the cytoplasm and nucleus.
    Science. 1985 Nov 15;230(4727):770-6 PMID: 2997917
  27. A new technique for the assay of infectivity of human adenovirus 5 DNA.
    Virology. 1973 Apr;52(2):456-67 PMID: 4705382
  28. DNA sequencing with chain-terminating inhibitors.
    Proc Natl Acad Sci U S A. 1977 Dec;74(12):5463-7 PMID: 271968
  29. The role of guanosine 5'-triphosphate in polypeptide chain elongation.
    Biochim Biophys Acta. 1978 Sep 21;505(1):95-127 PMID: 361078
  30. The amino acid sequence of elongation factor Tu of Escherichia coli. The complete sequence.
    J Biol Chem. 1981 Aug 10;256(15):8102-9 PMID: 7021545
  31. Transformation of mammalian cells to antibiotic resistance with a bacterial gene under control of the SV40 early region promoter.
    J Mol Appl Genet. 1982;1(4):327-41 PMID: 6286831
  32. The Rous sarcoma virus long terminal repeat is a strong promoter when introduced into a variety of eukaryotic cells by DNA-mediated transfection.
    Proc Natl Acad Sci U S A. 1982 Nov;79(22):6777-81 PMID: 6294651
  33. Complete nucleotide sequences of the T24 human bladder carcinoma oncogene and its normal homologue.
    Nature. 1983 Mar 3;302(5903):33-7 PMID: 6298635
  34. Splice junctions: association with variation in protein structure.
    Science. 1983 Jun 10;220(4602):1125-9 PMID: 6344214
  35. Nucleotide sequence of the Rasheed rat sarcoma virus oncogene: new mutations.
    Science. 1983 Jul 8;221(4606):155-7 PMID: 6344220
  36. Structure of the Ki-ras gene of the human lung carcinoma cell line Calu-1.
    Nature. 1983 Aug 11-17;304(5926):497-500 PMID: 6308465
  37. Structure and organization of the human Ki-ras proto-oncogene and a related processed pseudogene.
    Nature. 1983 Aug 11-17;304(5926):501-6 PMID: 6308466
  38. Transforming p21 ras protein: flexibility in the major variable region linking the catalytic and membrane-anchoring domains.
    EMBO J. 1985 Nov;4(11):2893-6 PMID: 2998761
  39. Mutant ras-encoded proteins with altered nucleotide binding exert dominant biological effects.
    Proc Natl Acad Sci U S A. 1986 Feb;83(4):952-6 PMID: 3513168
  40. Expression in rat fibroblasts of a human transforming growth factor-alpha cDNA results in transformation.
    Cell. 1986 Jul 18;46(2):301-9 PMID: 3459590
  41. GTP-binding proteins. One molecular machine can transduce diverse signals.
    Nature. 1986 Jun 26-Jul 2;321(6073):814-6 PMID: 3088455
  42. Identification of effector residues and a neutralizing epitope of Ha-ras-encoded p21.
    Proc Natl Acad Sci U S A. 1986 Jul;83(13):4725-9 PMID: 2425352
  43. The oncogenic activation of human p21ras by a novel mechanism.
    Science. 1986 Aug 8;233(4764):649-52 PMID: 3487832
  44. The ral gene: a new ras related gene isolated by the use of a synthetic probe.
    EMBO J. 1986 Sep;5(9):2203-8 PMID: 3023062
  45. Mutational analysis of a ras catalytic domain.
    Mol Cell Biol. 1986 Jul;6(7):2646-54 PMID: 3023943
  46. Structure of the human and murine R-ras genes, novel genes closely related to ras proto-oncogenes.
    Cell. 1987 Jan 16;48(1):137-46 PMID: 3098437
  47. Structure and activation of the human N-ras gene.
    Cell. 1983 Sep;34(2):581-6 PMID: 6616621
Article Info
Journal
Proceedings of the National Academy of Sciences of the United States of America
Abbr.
Proc Natl Acad Sci U S A
ISSN
0027-8424
DOI
10.1073/pnas.85.4.1015
Published
1988-02-00
Pages
1015-9
Language
English
Region
United States
NLM ID
7505876
PMCID
PMC279691
Subset
IM
External Links
PubMed source DOI: 10.1073/pnas.85.4.1015 PMC Full Text View journal details
Analysis Services
Analysis Services

Contact

No. 2 Wenbo Road, Zhangqiu District, Jinan, Shandong

Qilu Normal University · Genelibs Bioinformatics Lab

750 Shunhua Rd, Jinan

2F, Bldg F, University Science Park

Tel: 0531-88819269

WeChat Official Account

Follow our WeChat subscription account for real-time updates and the latest in medical and biological research.


Business Email

E-mail: [email protected]

Copyright © 2026.Genelibs Jinan Honghui Biotechnology Co., Ltd. 鲁ICP备20007159号-2