Sequences capable of restoring poly(A) site function define two distinct downstream elements.

M. A. McDevitt, R. P. Hart, W. W. Wong, J. R. Nevins

Research output: Contribution to journalArticle

Abstract

Several recent studies have shown that a functional poly(A) site consists of both an AAUAAA element as well as sequences downstream of the cleavage site. Two downstream regions were analyzed in an attempt to accurately locate and define the critical sequences. Chemically synthesized oligonucleotides of sequence from the early SV40 and the adenovirus E2A poly(A) sites were able to restore efficient cleavage to a deleted SV40 poly(A) site. Inversion of the sequence completely abolished poly(A) site function. A series of base substitution mutants were generated in each downstream sequence. Certain single base changes drastically altered poly(A) site function. Thus, it is concluded that a defined downstream sequence of limited complexity is important for efficient processing of the primary transcript at the poly(A) site. The position of the downstream elements relative to the AAUAAA and cleavage site was found to be critical since moving either the E2 element or the SV40 element an additional 40 nucleotides downstream abolished function. There were differences, however, in the effect of spacing on the function of the two elements. This observation, along with the fact that the two sequences are clearly different, indicates that there are at least two distinct genetic elements that direct efficient cleavage at the poly(A) site.

Original languageEnglish (US)
Pages (from-to)2907-2913
Number of pages7
JournalThe EMBO journal
Volume5
Issue number11
DOIs
StatePublished - Nov 1986

ASJC Scopus subject areas

  • Neuroscience(all)
  • Molecular Biology
  • Biochemistry, Genetics and Molecular Biology(all)
  • Immunology and Microbiology(all)

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