ArticleNucleic acids research2002
The role of template-primer in protection of reverse transcriptase from thermal inactivation.
Article in Nucleic acids research, 2002. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 31 papers.
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Who cites it
31 citing papers in PubMed, 88 citations in OpenAlex.
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- Applications of Quantitative PCR (qPCR) in Studies of Virus-Host Interactions.Methods in molecular biology (Clifton, N.J.) · 2024Article
- A Novel Thermostable and Processive Reverse Transcriptase from a Group II Intron ofBiomolecules · 2023Article
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- CRISPR/Cas9 therapeutics: progress and prospects.Signal transduction and targeted therapy · 2023Review
- Generation of an Avian Myeloblastosis Virus (AMV) Reverse Transcriptase Prime Editor.Advances in experimental medicine and biology · 2023Article
- Comparison of Reverse Transcriptase (RT) Activities of Various M-MuLV RTs for RT-LAMP Assays.Biology · 2022Article
- A flexible split prime editor using truncated reverse transcriptase improves dual-AAV delivery in mouse liver.Molecular therapy : the journal of the American Society of Gene Therapy · 2022Article
- Advances in single-cell sequencing technology in the field of hepatocellular carcinoma.Frontiers in genetics · 2022Review
- Targeted insertion of large genetic payloads using cas directed LINE-1 reverse transcriptase.Scientific reports · 2021Article
- Compendium of Plant-Specific CRISPR Vectors and Their Technical Advantages.Life (Basel, Switzerland) · 2021Review
- M-MuLV reverse transcriptase: Selected properties and improved mutants.Computational and structural biotechnology journal · 2021Review
- Alteration of enzymes and their application to nucleic acid amplification (Review).International journal of molecular medicine · 2020Review
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Authors and funding
7 authors at 1 institution in 1 country.
Funding
No grant is acknowledged in the PubMed record.
Abstract
We compared the thermal stabilities of wild-type recombinant avian myeloblastosis virus (AMV) and Moloney murine leukemia virus (M-MLV) reverse transcriptase (RT) with those of mutants of the recombinant enzymes lacking RNase H activity. They differed in resistance to thermal inactivation at elevated temperatures in the presence of an RNA/DNA template-primer. RNase H-minus RTs retained the ability to efficiently synthesize cDNA at much higher temperatures. We show that the structure of the template-primer has a critical bearing on protection of RT from thermal inactivation. RT RNase H activity rapidly alters the structure of the template-primer to forms less tightly bound by RT and thus less able to protect the enzyme at elevated temperatures. We also found that when comparing wild-type or mutant AMV RT with the respective M-MLV RT, the avian enzymes retained more DNA synthetic activity at elevated temperatures than murine RTs. Enzyme, template-primer interaction again played the most significant role in producing these differences. AMV RT binds much tighter to template- primer and has a much greater tendency to remain bound during cDNA synthesis than M-MLV RT and therefore is better protected from heat inactivation.
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