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Molecular Cloning and Characterization of GPA1, a G Protein {alpha} Subunit Gene from Arabidopsis thaliana

Ma, Hong and Yanofsky, Martin F. and Meyerowitz, Eliott M. (1990) Molecular Cloning and Characterization of GPA1, a G Protein {alpha} Subunit Gene from Arabidopsis thaliana. Proceedings of the National Academy of Sciences of the United States of America, 87 (10). pp. 3821-3825. ISSN 0027-8424. http://resolver.caltech.edu/CaltechAUTHORS:MAHpnas90

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Abstract

We have isolated a gene coding for a G protein {alpha} subunit from the flowering plant Arabidopsis thaliana. This gene, named GPA1, was isolated by using a DNA probe generated by polymerase chain reaction based on protein sequences from mammalian and yeast G protein {alpha} subunits. The sequences of genomic and cDNA clones indicate that GPA1 has 14 exons, and the deduced amino acid sequence shows that the GPA1 gene product (GP{alpha}1) has 383 amino acid residues (44,582 Da). The GP{alpha}1 protein exhibits similarity to all known G protein {alpha} subunits--36% of its amino acids are identical and 73% are similar (identical and conservative changes) to mammalian inhibitory guanine nucleotide-binding regulatory factor {alpha} subunits and transducins. Furthermore, the GP{alpha}1 protein has all of the consensus regions for a GTP-binding protein. The GPA1-encoded mRNA of 1.55 kilobases is most abundant in vegetative plant tissues, as determined by RNA blot analysis. Restriction fragment length polymorphism mapping experiments show that GPA1 is {approx} 1.2 centimorgans from the visible marker er on chromosome 2.


Item Type:Article
Additional Information:Copyright © 1990 by National Academy of Sciences Communicated by Melvin I. Simon, March 8, 1990. The sequence reported in this paper has been deposited in the GenBank data base (accession no. M32887). We thank M. Simon, M. Strathmann, M. Lochrie, and T. Wilkie for gifts of oligonucleotides, advice, and helpful discussions; A. van der Bliek for help with PCR experiments; J. Bowman for the cDNA library and RFLP DNA blots; G. Drews for helping with the RNA blot; S. Kempin for computer analysis of the RFLP mapping data; S. Kempin and Y. Hu for excellent technical assistance; and T. Stearns for help with the protein sequence data base search. We thank R. Bourret, L. Brockman, C. Chang, G. Drews, M. Lochrie, M. Simon, M. Strathmann, A. van der Bliek, D. Weigel, and T. Wilkie for critical reading of this manuscript. This work was supported by a grant from the National Science Foundation (DCB 8703439) and a grant from the Lucille P. Markey Charitable Trust to E.M.M. H.M. was supported by a postdoctoral fellowship from the Helen Hay Whitney Foundation, and M.F.Y. was supported by a postdoctoral fellowship in plant biology from the National Science Foundation (DCB 8608456). The publication costs of this article were defrayed in part by page charge payment. This article must therefore be hereby marked "advertisement" in accordance with 18 U.S.C. §1734 solely to indicate this fact.
Subject Keywords:polymerase chain reaction, GTP-binding protein, [alpha] subunit, restriction fragment length polymorphism mapping
Record Number:CaltechAUTHORS:MAHpnas90
Persistent URL:http://resolver.caltech.edu/CaltechAUTHORS:MAHpnas90
Alternative URL:http://www.pnas.org/cgi/content/abstract/87/10/3821
Usage Policy:No commercial reproduction, distribution, display or performance rights in this work are provided.
ID Code:662
Collection:CaltechAUTHORS
Deposited By: Archive Administrator
Deposited On:12 Sep 2005
Last Modified:26 Dec 2012 08:40

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