Sequence requirements for branch formation in a group ll self-splicing lntron

Rachel Altura, Brian Rymond, Bertrand Seraphin, Michael Rosbash

Research output: Contribution to journalArticlepeer-review

17 Scopus citations

Abstract

Evidence is presented for the existence of a specific intronintron interaction, necessary for the formation of the branched product in the self-splicing reaction of a group II yeast mitochondrial intron. Trans-splicing reactions involving two RNA molecules (5'exon with covalently linked regions of intron and intron with covalently linked 3'exon) show that the presence of portions of intron domain I on the 51 molecule is necessary for the formation of branched products which are not seen with shorter 51 molecules. Modification/interference reactions show regions necessary for branch-formation and support a major role for specific regions of intron domain I. Further experiments, utilizing a truncated 31 molecule that is missing the conserved branchpoint nucleotide, indicate that domain VI may be required for a successful domain I interaction. A model for the formation of a proper branched structure includes implications for both cis and trans configurations.

Original languageEnglish
Pages (from-to)335-354
Number of pages20
JournalNucleic Acids Research
Volume17
Issue number1
DOIs
StatePublished - Jan 11 1989

Bibliographical note

Funding Information:
ACKNOWLEDGEMENTS We thank Michael Green for his generous gift of debranching extract and Claudio Pikielny for his comments on the manuscript. We especially thank all members of the Rosbash laboratory for their thoughtful suggestions during the course of this study and, in the preparation of the manuscript, specifically, Hildur V. Colot for her writing suggestions, and Tobie Tishman for her assistance in compiling this manuscript. Most of this work was submitted by R.A. to the Brandeis University Department of Biology in partial fulfillment of the requirements for an undergraduate honors degree. This work was supported by National Institutes of Health grant GM23549.

ASJC Scopus subject areas

  • Genetics

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