HOME HELP FEEDBACK SUBSCRIPTIONS ARCHIVE SEARCH TABLE OF CONTENTS		QUICK SEARCH:   [advanced] Author: Keyword(s): Year:  Vol:  Page:

J. Biol. Chem., Vol. 279, Issue 27, 28539-28552, July 2, 2004

This Article Full Text Full Text (PDF) All Versions of this Article: 279/27/28539    most recent M310138200v1 Submit a Letter to Editor Alert me when this article is cited Alert me when eLetters are posted Alert me if a correction is posted Citation Map Services Email this article to a friend Similar articles in this journal Similar articles in PubMed Alert me to new issues of the journal Download to citation manager Request Permissions Citing Articles Citing Articles via HighWire Citing Articles via Google Scholar Google Scholar Articles by Majee, M. Articles by Majumder, A. L. Search for Related Content PubMed PubMed Citation Articles by Majee, M. Articles by Majumder, A. L. Social Bookmarking                      What's this?

A Novel Salt-tolerant L-myo-Inositol-1-phosphate Synthase from Porteresia coarctata (Roxb.) Tateoka, a Halophytic Wild Rice

MOLECULAR CLONING, BACTERIAL OVEREXPRESSION, CHARACTERIZATION, AND FUNCTIONAL INTROGRESSION INTO TOBACCO-CONFERRING SALT TOLERANCE PHENOTYPE^*

Manoj Majee, Susmita Maitra, Krishnarup Ghosh Dastidar, Sitakanta Pattnaik, Anirban Chatterjee, Nitai C. Hait¶, Kali Pada Das||, and Arun Lahiri Majumder**

From the Plant Molecular and Cellular Genetics and the ^||Department of Chemistry, Bose Institute (Centenary Building), P-1/12, C I T Scheme VII M, Kolkata 700054, India

L-myo-Inositol-1-phosphate synthase (EC 5.5.1.4, MIPS), an evolutionarily conserved enzyme protein, catalyzes the synthesis of inositol, which is implicated in a number of metabolic reactions in the biological kingdom. Here we report on the isolation of the gene (PINO1) for a novel salt-tolerant MIPS from the wild halophytic rice, Porteresia coarctata (Roxb.) Tateoka. Identity of the PINO1 gene was confirmed by functional complementation in a yeast inositol auxotrophic strain. Comparison of the nucleotide and deduced amino acid sequences of PINO1 with that of the homologous gene from Oryza sativa L. (RINO1) revealed distinct differences in a stretch of 37 amino acids, between amino acids 174 and 210. Purified bacterially expressed PINO1 protein demonstrated a salt-tolerant character in vitro compared with the salt-sensitive RINO1 protein as with those purified from the native source or an expressed salt-sensitive mutant PINO1 protein wherein amino acids 174–210 have been deleted. Analysis of the salt effect on oligomerization and tryptophan fluorescence of the RINO1 and PINO1 proteins revealed that the structure of PINO1 protein is stable toward salt environment. Furthermore, introgression of PINO1 rendered transgenic tobacco plants capable of growth in 200–300 mM NaCl with retention of 40–80% of the photosynthetic competence with concomitant increased inositol production compared with unstressed control. MIPS protein isolated from PINO1 transgenics showed salt-tolerant property in vitro confirming functional expression in planta of the PINO1 gene. To our knowledge, this is the first report of a salt-tolerant MIPS from any source.

Received for publication, September 12, 2003 , and in revised form, February 23, 2004.

The nucleotide sequence(s) reported in this paper has been submitted to the GenBank^TM/EBI Data Bank with accession number(s) AF412340.

^* This work was supported by research grants from the Department of Biotechnology, the Department of Science and Technology, and the Council of Scientific and Industrial Research, Government of India. The costs of publication of this article were defrayed in part by the payment of page charges. This article must therefore be hereby marked "advertisement"in accordance with 18 U.S.C. Section 1734 solely to indicate this fact.

Supported by a Fellowship of the Department of Biotechnology Postdoctoral Training Programme. Present address: Kentucky Tobacco Research and Development Center, University of Kentucky, Lexington, KY 40546-0236.

^¶ Present address: Dept. of Biochemistry, Medical College of Virginia, Virginia Commonwealth University, Richmond, VA 23298-0614.

^** To whom correspondence should be addressed: Plant Molecular and Cellular Genetics, Bose Institute (Centenary Building), P-1/12, C I T Scheme VII M, Kolkata 700054, India. Tel.: 91-33-2337-9544; Fax: 91-33-2334-3886; E-mail: lahiri{at}bic.boseinst.ernet.in.

CiteULike    Complore    Connotea    Del.icio.us    Digg    Reddit    Technorati    What's this?

This article has been cited by other articles:

				E. F. M. Abreu and F. J. L. Aragao Isolation and Characterization of a myo-inositol-1-phosphate Synthase Gene from Yellow Passion Fruit (Passiflora edulis f. flavicarpa) Expressed During Seed Development and Environmental Stress Ann. Bot., February 1, 2007; 99(2): 285 - 292. [Abstract] [Full Text] [PDF]

				K. Ghosh Dastidar, S. Maitra, L. Goswami, D. Roy, K. P. Das, and A. L. Majumder An Insight into the Molecular Basis of Salt Tolerance of L-myo-Inositol 1-P Synthase (PcINO1) from Porteresia coarctata (Roxb.) Tateoka, a Halophytic Wild Rice Plant Physiology, April 1, 2006; 140(4): 1279 - 1296. [Abstract] [Full Text] [PDF]