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PLANT PHYSIOLOGY , Vol 102, Issue 1 227-232, Copyright © 1993 by American Society of Plant Biologists

MOLECULAR BIOLOGY AND GENE REGULATION

Molecular Basis of the Ribulose-1,5-Bisphosphate Carboxylase/Oxygenase Activase Mutation in Arabidopsis thaliana Is a Guanine-to-Adenine Transition at the 5[prime]-Splice Junction of Intron 3

B. M. Orozco, C. R. McClung, J. M. Werneke and W. L. Ogren
Department of Plant Biology, University of Illinois at Urbana-Champaign (B.M.O., J.M.W., W.L.O); Department of Biological Sciences, Dartmouth College, Hanover, New Hampshire (C.R.M.); and Photosynthesis Research Unit, Agricultural Research Service, United States Department of Agriculture, Urbana, Illinois 61801-3838 (W.L.O.)

Analysis of the ribulose-1,5-bisphosphate carboxylase/oxygenase (Rubisco) activase gene and gene products from Arabidopsis thaliana wild-type plants and the Rubisco activase-deficient mutant strain showed that the rca mutation caused GT to be changed to AT at the 5[prime]-splice junction of intron 3 in the six-intron pre-mRNA. Northern blot analysis, genomic and cDNA sequencing, and primer extension analysis indicated that the mutation causes inefficient and incomplete splicing of the pre-mRNA, resulting in the accumulation of three aberrant mRNAs. One mutant mRNA was identical with wild-type mRNA except that it included intron 3, a second mRNA comprised intron 3 and exons 4 through 7, and the third mRNA contained exons 1 through 3. The G-to-A transition is consistent with the known mechanism of mutagenesis by ethyl methanesulfonate, the mutagen used to create the Rubisco activase-deficient strain.


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