ArticleThe Biochemical journal2000
Expression of an active form of recombinant Ty1 reverse transcriptase in Escherichia coli: a fusion protein containing the C-terminal region of the Ty1 integrase linked to the reverse transcriptase-RNase H domain exhibits polymerase and RNase H activities.
Article in The Biochemical journal, 2000. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 21 papers.
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Who cites it
21 citing papers in PubMed, 40 citations in OpenAlex.
- A proteomic screen of Ty1 integrase partners identifies the protein kinase CK2 as a regulator of Ty1 retrotransposition.Mobile DNA · 2022Article
- Ty1 integrase is composed of an active N-terminal domain and a large disordered C-terminal module dispensable for its activity in vitro.The Journal of biological chemistry · 2021Article
- The Ty1 LTR-retrotransposon of budding yeast,Microbiology spectrum · 2015Article
- Retrotransposon Ty1 RNA contains a 5'-terminal long-range pseudoknot required for efficient reverse transcription.RNA (New York, N.Y.) · 2013Article
- Effect of reverse transcriptase inhibitors on LINE-1 and Ty1 reverse transcriptase activities and on LINE-1 retrotransposition.BMC biochemistry · 2011Article
- GeneDesign 3.0 is an updated synthetic biology toolkit.Nucleic acids research · 2010Article
- Posttranslational interference of Ty1 retrotransposition by antisense RNAs.Proceedings of the National Academy of Sciences of the United States of America · 2009Article
- The diversity of retrotransposons and the properties of their reverse transcriptases.Virus research · 2008Review
- Article
- Ty1 reverse transcriptase does not read through the proposed 2',5'-branched retrotransposition intermediate in vitro.RNA (New York, N.Y.) · 2007Article
- Mn2+ suppressor mutations and biochemical communication between Ty1 reverse transcriptase and RNase H domains.Journal of virology · 2007Article
- Cooperation between reverse transcriptase and integrase during reverse transcription and formation of the preintegrative complex of Ty1.Eukaryotic cell · 2006Article
- Role of integrase in reverse transcription of the Saccharomyces cerevisiae retrotransposon Ty1.Eukaryotic cell · 2005Article
- Functional roles of carboxylate residues comprising the DNA polymerase active site triad of Ty3 reverse transcriptase.Nucleic acids research · 2005Article
- A group II intron-type open reading frame from the thermophile Bacillus (Geobacillus) stearothermophilus encodes a heat-stable reverse transcriptase.Applied and environmental microbiology · 2004Article
- A 5'-3' long-range interaction in Ty1 RNA controls its reverse transcription and retrotransposition.The EMBO journal · 2002Article
- Ty1 defect in proteolysis at high temperature.Journal of virology · 2002Article
- Targeting of the yeast Ty5 retrotransposon to silent chromatin is mediated by interactions between integrase and Sir4p.Molecular and cellular biology · 2001Article
- The protease and reverse transcriptase of the tobacco LTR retrotransposon Tnt1 are enzymatically active when expressed in Escherichia coli.Plant molecular biology · 2001Article
- A Ty1 reverse transcriptase active-site aspartate mutation blocks transposition but not polymerization.Journal of virology · 2001Article
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Authors and funding
3 authors at 2 institutions in 1 country.
Funding
No grant is acknowledged in the PubMed record.
Abstract
Replication of the Saccharomyces cerevisiae Ty1 retrotransposon requires a reverse transcriptase capable of synthesizing Ty1 DNA. The first description of an active form of a recombinant Ty1 enzyme with polymerase and RNase H activities is reported here. The Ty1 enzyme was expressed as a hexahistidine-tagged fusion protein in Escherichia coli to facilitate purification of the recombinant protein by metal-chelate chromatography. Catalytic activity of the recombinant protein was detected only when amino acid residues encoded by the integrase gene were added to the N-terminus of the reverse transcriptase-RNase H domain. This suggests that the integrase domain could play a role in proper folding of reverse transcriptase. Several biochemical properties of the Ty1 enzyme were analysed, including the effect of MgCl(2), NaCl, temperature and of the chain terminator dideoxy GTP on its polymerase activity. RNase H activity was examined by monitoring the cleavage of a RNA-DNA template-primer. Our results suggest that the distance between the RNase H and polymerase active sites corresponds to the length of a 14-nucleotide RNA-DNA heteroduplex. The recombinant protein produced in E. coli should be useful for further biochemical and structural analyses and for a better understanding of the role of integrase in the activation of reverse transcriptase.
Indexed as
Identifiers
10816427PMC1221071W1976762486What OpenQuestion holds
Registered trials
Read under generation 80e0d062 · epoch 390. Bibliography from PubMed, PubMed Central and OpenAlex; grants from NIH RePORTER; trial links from ClinicalTrials.gov; estimates, votes and beliefs from the OpenQuestion graph.