Aug 5, 1989

Plus-strand priming by Moloney murine leukemia virus. The sequence features important for cleavage by RNase H

Journal of Molecular Biology
A J Rattray, James J Champoux

Abstract

The reverse transcriptase-associated RNase H activity is responsible for producing the plus-strand RNA primer during reverse transcription. The major plus-strand initiation site is located within a highly conserved polypurine tract (PPT), and initiation of DNA replication at this site is necessary for proper formation of the viral long terminal repeats (LTRs). We present here a compilation of PPT sequences from an evolutionarily diverse group of retroviruses and retrotransposons, which reveals that there is a high degree of sequence conservation at this site. Furthermore, we found previously that secondary plus-strand origins, identified in vitro, also show strong similarity to the PPT. Taken together, these data suggest that RNase H recognizes a specific sequence at the PPT as a signal to cleave the RNA at a precise location, producing a primer for the initiation of plus-DNA strands. We have analyzed the RNase H recognition sequence by producing a large number of single and double mutations within the PPT. Our findings suggest that no single residue in the +5 to -6 region (where the cleavage occurs between -1 and +1) is essential; mutations at these positions introduced heterogeneity at the cleavage site, but cleavage is still...Continue Reading

Mentioned in this Paper

Long Terminal Repeat
DNA, Viral
Transcription Initiation
Cytokinesis of the Fertilized Ovum
Retrotransposons
Oligonucleotide Primers
Cytokinesis
Moloney Leukemia Virus
Genomic Hybridization
DNA Replication

About this Paper

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