reverse transcriptase

reverse transcriptase
Thomas Splettstoesser (www.scistyle.com) · CC BY-SA 3.0 · source
Name	Reverse transcriptase
Ec number	2.7.7.49
Cofactors	Mg2+, Mn2+

reverse transcriptase

Reverse transcriptase is an enzyme that synthesizes complementary DNA from an RNA template. Discovered in the context of Howard Temin's and David Baltimore's independent work, it challenged central dogma concepts and influenced research across National Institutes of Health, Cold Spring Harbor Laboratory, Rockefeller University, and Institut Pasteur. The enzyme underpins technologies developed by entities such as Genentech, Applied Biosystems, and Thermo Fisher Scientific and has had major impact on fields shaped at institutions like Massachusetts Institute of Technology and Stanford University.

Introduction

Reverse transcriptase catalyzes RNA-dependent DNA polymerization and possesses distinct activities including RNA-dependent DNA polymerase, DNA-dependent DNA polymerase, and RNase H in many forms. Key historical milestones involved work recognized by the Nobel Prize in Physiology or Medicine awarded to Howard Temin and David Baltimore and the subsequent debates at gatherings like conferences organized by Cold Spring Harbor Laboratory and publications in journals associated with Nature Publishing Group and Cell Press.

Structure and Mechanism

Structurally, reverse transcriptases are heterodimeric or monomeric proteins with domains analogous to a polymerase "hand" comprising fingers, palm, and thumb substructures; many features were elucidated by researchers at Max Planck Society-affiliated laboratories and through crystallography at facilities such as Brookhaven National Laboratory and European Synchrotron Radiation Facility. The active site coordinates divalent cations like magnesium and manganese and interacts with nucleoside triphosphates in a manner similar to other polymerases studied at Cold Spring Harbor Laboratory and Salk Institute. RNase H domains cleave RNA in RNA–DNA hybrids, a functional motif explored by teams at Johns Hopkins University and University of Cambridge. High-resolution structures informed by work at Lawrence Berkeley National Laboratory and solved by groups sponsored by foundations like the Howard Hughes Medical Institute revealed conformational transitions during translocation and strand displacement analogous to mechanisms characterized in studies at European Molecular Biology Laboratory and University of Oxford.

Biological Roles and Occurrence

Reverse transcriptases occur in retroviruses such as Human immunodeficiency virus, endogenous retroelements including LINE-1, and in retrotransposons characterized in genomes sequenced by consortia like the Human Genome Project and projects at Wellcome Trust Sanger Institute. They are produced by retroviruses studied in virology labs at Centers for Disease Control and Prevention and by mobile genetic elements analyzed by researchers at National Center for Biotechnology Information and European Bioinformatics Institute. Reverse transcription mediates life-cycle steps in pathogens investigated by teams at World Health Organization-associated networks and affects genome architecture in organisms from Saccharomyces cerevisiae to humans, with evolutionary consequences assessed in comparative genomics efforts led by Broad Institute and J. Craig Venter Institute.

Applications and Uses

Reverse transcriptase underlies methods such as reverse transcription polymerase chain reaction developed by companies including Roche and platforms from Bio-Rad Laboratories and Illumina. It enabled cDNA cloning workflows employed in laboratories at Harvard University and Yale University and contributed to transcriptomics projects undertaken by consortia like the ENCODE Project Consortium and initiatives at European Molecular Biology Laboratory. Clinical diagnostics using RT-PCR were scaled by public health agencies such as Centers for Disease Control and Prevention and diagnostic divisions at Roche Diagnostics and informed large-scale surveillance by networks coordinated with World Health Organization. Reverse transcriptase–based methods support technologies commercialized by Agilent Technologies and academic spinouts from institutions like University of California, Berkeley.

Inhibitors and Clinical Relevance

Reverse transcriptase inhibitors, both nucleoside and non-nucleoside classes, are cornerstone therapeutics in treatment regimens developed by pharmaceutical companies including Gilead Sciences, Merck & Co., and GlaxoSmithKline. Antiretroviral therapy guidelines issued by bodies such as World Health Organization and American Academy of Pediatrics incorporate drugs that target reverse transcriptase to manage infections like AIDS caused by Human immunodeficiency virus. Clinical trials overseen by agencies including the Food and Drug Administration and funded by institutions like the Wellcome Trust and Bill & Melinda Gates Foundation evaluated combination therapies and resistance profiles monitored through surveillance programs run by European Medicines Agency and national public health institutes.

Evolutionary Significance and Diversity

Reverse transcriptase has shaped genome evolution via retrotransposition events cataloged by genome projects at the Human Genome Project and analyses by the 1000 Genomes Project and ENCODE Project Consortium. Variants of reverse transcriptase are found across domains of life in retroelements studied by teams at Cold Spring Harbor Laboratory and in retroviral lineages traced through phylogenetic work conducted by researchers at Smithsonian Institution and university groups worldwide. The enzyme's discovery influenced theoretical frameworks discussed at meetings organized by National Academy of Sciences and in monographs from presses such as Oxford University Press and Cambridge University Press that examine molecular evolution and the history of biology.

Category:Enzymes