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PMID: 7527135 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't

Yeast Ty1 retrotransposon: the minus-strand primer binding site and a cis-acting domain of the Ty1 RNA are both important for packaging of primer tRNA inside virus-like particles.

Nucleic acids research ·Vol. 22 ·No. 22 ·1994-11-11 ·Pages 4560-5

Wilhelm M, Wilhelm FX, Keith G, Agoutin B, Heyman T

Abstract

Reverse transcription of the yeast retrotransposon Ty1 is primed by the cytoplasmic initiator methionine tRNA (tRNA(iMet)). The primer tRNA(iMet) is packaged inside virus-like particles (VLPs) and binds to a 10 nucleotides minus-strand primer binding site, the (-)PBS, complementary to its 3' acceptor stem. We have found that three short sequences of the Ty1 RNA (box 1, box 2.1 and box 2.2) located 3' to the (-)PBS are complementary to other regions of the primer tRNA(iMet) (T psi C and DHU stems and loops). Reconstitution of reverse transcription in vitro with T7 transcribed Ty1 RNA species and tRNA(iMet) purified from yeast cells shows that the boxes do not affect the efficiency of reverse transcription. Thus the role of the boxes on packaging of the primer tRNA(iMet) into the VLPs was investigated by analysing the level of tRNA(iMet) packaged into mutant VLPs. Specific nucleotide changes in the (-)PBS or in the boxes that do not change the protein coding sequence but disrupt the complementarity with the primer tRNA(iMet) within the VLPs. We propose that base pairing between the primer tRNA(iMet) and the Ty1 RNA is of major importance for tRNA(iMet) packaging into the VLPs. Moreover the intactness of the boxes is essential for retrotransposition as shown by the transposition defect of a Ty1 element harboring an intact (-)PBS and mutated boxes.

MeSH Terms
Amino Acid Sequence Base Sequence Binding Sites Cloning, Molecular Molecular Sequence Data Mutation/physiology RNA/genetics RNA, Fungal/genetics RNA, Transfer, Met/genetics Retroelements/genetics,physiology Retroviridae/genetics Saccharomyces cerevisiae/genetics Transcription, Genetic
Chemicals
RNA primers RNA, Fungal RNA, Transfer, Met Retroelements RNA
Authors & Affiliations
5 authors, click to expand affiliations / ORCID
Wilhelm M
Unité Propre de Recherche 9002 du Centre National de la Recherche Scientifique, Institut de Biologie Moléculaire et Cellulaire, France.
Wilhelm F X
Keith G
Agoutin B
Heyman T
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Article Info
Journal
Nucleic acids research
Abbr.
Nucleic Acids Res
ISSN
0305-1048
DOI
10.1093/nar/22.22.4560
Published
1994-11-11
Pages
4560-5
Language
English
Region
England
NLM ID
0411011
PMCID
PMC308501
Subset
IM
External Links
PubMed source DOI: 10.1093/nar/22.22.4560 PMC Full Text View journal details
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