Isolation of Schizosaccharomyces pombe isopentenyl diphosphate isomerase cDNA clones by complementation and synthesis of the enzyme in Escherichia coli

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Publication Type Journal Article
School or College College of Science
Department Chemistry
Creator Poulter, Charles Dale
Other Author Hahn, Frederick M.
Title Isolation of Schizosaccharomyces pombe isopentenyl diphosphate isomerase cDNA clones by complementation and synthesis of the enzyme in Escherichia coli
Date 1995-05-12
Description Isopentenyl diphosphate (IPP) isomerase catalyzes an essential activation step in the isoprene biosynthetic pathway. The Saccharomyces cerevisiae gene for IPP isomerase, IDI1, was recently isolated and characterized (Anderson, M. S., Muehlbacher, M., Street, I. P., Proffitt, J., and Poulter, C. D. (1989) J. Biol. Chem. 264, 19169-19175), and the wild-type gene, IDI1, was disrupted with a LEU2 marker to create a diploid yeast strain heterozygous for the idi1::leu2 disruption, which revealed that IDI1 was an essential single-copy gene (Mayer, M.P., Hahn, F. M., Stillman, D. J., and Poulter, C. D. (1992) Yeast 8, 743-748). We now report the isolation of a cDNA clone from Schizosaccharomyces pombe by a plasmid shuffle-mediated complementation of the LEU2 disrupted yeast gene. The S. pombe clone encoded a 26,864-dalton polypeptide of 227 amino acids with a high degree of similarity to the S. cerevisiae IDI1 enzyme. S. pombe IPP isomerase contained the essential Cys and Glu catalytic residues identified in yeast isomerase (Street, I. P., Coffman, H. R., Baker, J., and Poulter, C. (1994) Biochemistry 33, 4212-4217) but was significantly smaller than the S. cerevisiae enzyme. The plasmid shuffle technique is an excellent procedure for screening expression libraries for IPP isomerase activity by complementation of the idi1 mutation.
Type Text
Publisher American Society for Biochemistry and Molecular Biology (ASBMB)
Volume 270
First Page 11298
Last Page 11303
Subject Carbon-Carbon; Biosynthesis; Genetics
Subject LCSH Biosynthesis; Genetics
Dissertation Institution University of Utah
Language eng
Bibliographic Citation Poulter, C. D. & Hahn, F. M. (1995). Isolation of Schizosaccharomyces pombe isopentenyl diphosphate isomerase cDNA clones by complementation and synthesis of the enzyme in Escherichia coli. Journal of Biological Chemistry. 270, 11298-303.
Rights Management (c)American Society for Biochemistry and Molecular Biology (ASBMB)
Format Medium application/pdf
Identifier ir-main,12095
ARK ark:/87278/s6h4294t
Setname ir_uspace
Date Created 2012-06-13
Date Modified 2013-01-31
ID 707220
Reference URL https://collections.lib.utah.edu/ark:/87278/s6h4294t
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