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Plant J. 1996 Mar;9(3):381-9.

Mapping of branchpoint nucleotides in mutant pre-mRNAs expressed in plant cells.

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1
Friedrich Miescher-Institut, Basel, Switzerland.

Abstract

Pre-mRNA encoding rubisco activase in the Arabidopsis thaliana mutant rca contains a GU to AU change at the 5' splice site of intron 3 and this mutation results in accumulation of splicing intermediates bearing an incompletely processed intron. It has been demonstrated that one of the intermediates contains intron 3 in the form of a lariat and the branchpoint nucleotide has been mapped to the A residue at position -32 forming part of the sequence UUGAU. Analysis of a similar GU to AU 5' splice site mutation, present in a synthetic pre-mRNA context expressed in transfected protoplasts of Nicotiana plumbaginifolia, also suggests formation of lariats with branching occurring at A-31. A small fraction (approximately 10%) of this mutant pre-mRNA also underwent the second step of splicing. In addition to the consensus AG, an AU dinucleotide was used as splicing acceptor.

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