Distinct Conformational States Underlie Pausing during Initiation of HIV-1 Reverse Transcription.

Journal of Molecular Biology
Kevin P LarsenElisabetta Viani Puglisi

Abstract

A hallmark of the initiation step of HIV-1 reverse transcription, in which viral RNA genome is converted into double-stranded DNA, is that it is slow and non-processive. Biochemical studies have identified specific sites along the viral RNA genomic template in which reverse transcriptase (RT) stalls. These stalling points, which occur after the addition of three and five template dNTPs, may serve as checkpoints to regulate the precise timing of HIV-1 reverse transcription following viral entry. Structural studies of reverse transcriptase initiation complexes (RTICs) have revealed unique conformations that may explain the slow rate of incorporation; however, questions remain about the temporal evolution of the complex and features that contribute to strong pausing during initiation. Here we present cryo-electron microscopy and single-molecule characterization of an RTIC after three rounds of dNTP incorporation (+3), the first major pausing point during reverse transcription initiation. Cryo-electron microscopy structures of a +3 extended RTIC reveal conformational heterogeneity within the RTIC core. Three distinct conformations were identified, two of which adopt unique, likely off-pathway, intermediates in the canonical polymer...Continue Reading

References

Jan 24, 1985·Nature·L RatnerK Baumeister
Jun 27, 1970·Nature·H M Temin, S Mizutani
Feb 14, 1995·Proceedings of the National Academy of Sciences of the United States of America·D W RodgersS C Harrison
Apr 4, 1998·Nucleic Acids Research·C IselR Marquet
Aug 8, 1998·The Journal of Biological Chemistry·C LiangM A Wainberg
Apr 15, 2000·The Journal of Biological Chemistry·J M LanchyR Marquet
Jun 1, 2001·The Journal of Biological Chemistry·N BeerensB Berkhout
Aug 27, 2002·The Journal of Biological Chemistry·Valerie GoldschmidtRoland Marquet
Jun 15, 2004·The Journal of Biological Chemistry·Valérie GoldschmidtRoland Marquet
Jul 21, 2004·Journal of Computational Chemistry·Eric F PettersenThomas E Ferrin
Dec 2, 2004·Acta Crystallographica. Section D, Biological Crystallography·Paul Emsley, Kevin Cowtan
Sep 27, 2005·Journal of Structural Biology·David N Mastronarde
Apr 25, 2007·Nucleic Acids Research·Ian W DavisDavid C Richardson
Aug 8, 2009·Nature·Joseph M WattsKevin M Weeks
Feb 4, 2010·Acta Crystallographica. Section D, Biological Crystallography·Paul D AdamsPeter H Zwart
Nov 26, 2010·Nature Structural & Molecular Biology·Shixin LiuXiaowei Zhuang
Jul 19, 2011·Journal of Molecular Biology·Elisabetta Viani Puglisi, Joseph D Puglisi
Jan 1, 2010·Viruses·Catherine IselRoland Marquet
Jan 24, 2012·Nature Structural & Molecular Biology·Kalyan DasEddy Arnold
Aug 30, 2012·Nature Methods·Caroline A SchneiderKevin W Eliceiri
Sep 25, 2012·Journal of Structural Biology·Sjors H W Scheres
Oct 3, 2012·Cold Spring Harbor Perspectives in Medicine·Wei-Shau Hu, Stephen H Hughes
Sep 17, 2013·Methods in Enzymology·Alexey PetrovJoseph D Puglisi
Dec 9, 2014·Journal of Structural Biology·Sjors H W Scheres
Nov 26, 2015·Journal of Structural Biology·Kai Zhang
Aug 31, 2016·Methods in Enzymology·S H W Scheres
Apr 27, 2018·Nature·Kevin P LarsenElisabetta Viani Puglisi
Jun 7, 2018·Acta Crystallographica. Section D, Structural Biology·Pavel V AfoninePaul D Adams
Sep 12, 2018·Journal of Molecular Biology·Aaron T CoeyElisabetta Viani Puglisi

❮ Previous
Next ❯

Citations

Sep 12, 2020·Current Opinion in Structural Biology·Miri KrupkinElisabetta Viani Puglisi

❮ Previous
Next ❯

Related Concepts

Trending Feeds

COVID-19

Coronaviruses encompass a large family of viruses that cause the common cold as well as more serious diseases, such as the ongoing outbreak of coronavirus disease 2019 (COVID-19; formally known as 2019-nCoV). Coronaviruses can spread from animals to humans; symptoms include fever, cough, shortness of breath, and breathing difficulties; in more severe cases, infection can lead to death. This feed covers recent research on COVID-19.

Blastomycosis

Blastomycosis fungal infections spread through inhaling Blastomyces dermatitidis spores. Discover the latest research on blastomycosis fungal infections here.

Nuclear Pore Complex in ALS/FTD

Alterations in nucleocytoplasmic transport, controlled by the nuclear pore complex, may be involved in the pathomechanism underlying multiple neurodegenerative diseases including Amyotrophic Lateral Sclerosis and Frontotemporal Dementia. Here is the latest research on the nuclear pore complex in ALS and FTD.

Applications of Molecular Barcoding

The concept of molecular barcoding is that each original DNA or RNA molecule is attached to a unique sequence barcode. Sequence reads having different barcodes represent different original molecules, while sequence reads having the same barcode are results of PCR duplication from one original molecule. Discover the latest research on molecular barcoding here.

Chronic Fatigue Syndrome

Chronic fatigue syndrome is a disease characterized by unexplained disabling fatigue; the pathology of which is incompletely understood. Discover the latest research on chronic fatigue syndrome here.

Evolution of Pluripotency

Pluripotency refers to the ability of a cell to develop into three primary germ cell layers of the embryo. This feed focuses on the mechanisms that underlie the evolution of pluripotency. Here is the latest research.

Position Effect Variegation

Position Effect Variagation occurs when a gene is inactivated due to its positioning near heterochromatic regions within a chromosome. Discover the latest research on Position Effect Variagation here.

STING Receptor Agonists

Stimulator of IFN genes (STING) are a group of transmembrane proteins that are involved in the induction of type I interferon that is important in the innate immune response. The stimulation of STING has been an active area of research in the treatment of cancer and infectious diseases. Here is the latest research on STING receptor agonists.

Microbicide

Microbicides are products that can be applied to vaginal or rectal mucosal surfaces with the goal of preventing, or at least significantly reducing, the transmission of sexually transmitted infections. Here is the latest research on microbicides.