A novel role for the 3' region of introns in pre-mRNA splicing of Saccharomyces cerevisiae.

B. C. Rymond, D. D. Torrey, M. Rosbash

Research output: Contribution to journalArticlepeer-review

18 Scopus citations


To investigate the importance of sequences between the yeast (Saccharomyces cerevisiae) branch point (TACTAAC box) and 3' splice site (AG), we generated a series of pre-mRNA substrates that differed in the length of RNA retained on the 3' side of the TACTAAC box. These pre-mRNAs were compared as substrates for the first step of in vitro splicing (5' cleavage and lariat formation) and in vitro spliceosome assembly (complex formation) in a whole-cell yeast extract. The results indicate that for rp51A pre-mRNA at least 29 nucleotides of RNA on the 3' side of the TACTAAC box are required for 5' cleavage and lariat formation, as smaller substrates fail to manifest any detectable cleavage or ligation events. Analysis of splicing complex assembly indicates that these smaller substrates undergo efficient yet incomplete complex formation; they are blocked at a late stage of spliceosome assembly, the complex I to complex II transition (Pikielny et al. 1986), a result which suggests that the failure to form lariats is due to a specific assembly defect. The lariat formation block (and assembly defect) can be relieved by the addition of ribohomopolymer "tails" to the 3' end of the shortened rp51A pre-mRNAs, and similar results were obtained with shortened actin pre-mRNAs. The results of this study indicate that this region of the pre-mRNA serves a specific function late in in vitro spliceosome assembly.

Original languageEnglish
Pages (from-to)238-246
Number of pages9
JournalGenes and Development
Issue number3
StatePublished - May 1987

ASJC Scopus subject areas

  • Genetics
  • Developmental Biology


Dive into the research topics of 'A novel role for the 3' region of introns in pre-mRNA splicing of Saccharomyces cerevisiae.'. Together they form a unique fingerprint.

Cite this