Mapping of nucleic acid binding in proteolytic domains of HIV-1 reverse transcriptase

Biochemistry ◽  
1993 ◽  
Vol 32 (29) ◽  
pp. 7466-7474 ◽  
Author(s):  
Amalendra Kumar ◽  
Hyeung Rak Kim ◽  
Robert W. Sobol ◽  
S. Patricia Becerra ◽  
Byeong Jae Lee ◽  
...  
Keyword(s):  
Nucleic Acid ◽  
Reverse Transcriptase ◽  
Nucleic Acid Binding ◽  
Hiv 1

scholarly journals A non-cleavable hexahistidine affinity tag at the carboxyl-terminus of the HIV-1 Pr55Gag polyprotein alters nucleic acid binding properties

2017 ◽  
Vol 130 ◽  
pp. 137-145 ◽  
Author(s):  
Maria C. Bewley ◽  
Lisa Reinhart ◽  
Matthew S. Stake ◽  
Shorena Nadaraia-Hoke ◽  
Leslie J. Parent ◽  
...  
Keyword(s):  
Nucleic Acid ◽  
Carboxyl Terminus ◽  
Nucleic Acid Binding ◽  
Affinity Tag ◽  
Binding Properties ◽  
Hiv 1

HIV-1 Nucleocapsid Traps Reverse Transcriptase on Nucleic Acid Substrates†

Biochemistry ◽  
2008 ◽  
Vol 47 (46) ◽  
pp. 12230-12240 ◽  
Author(s):  
Dina Grohmann ◽  
Julien Godet ◽  
Yves Mély ◽  
Jean-Luc Darlix ◽  
Tobias Restle
Keyword(s):  
Nucleic Acid ◽  
Reverse Transcriptase ◽  
Hiv 1

scholarly journals Two Coselected Distal Mutations in HIV-1 Reverse Transcriptase (RT) Alter Susceptibility to Nonnucleoside RT Inhibitors and Nucleoside Analogs

2019 ◽  
Vol 93 (11) ◽  
Author(s):  
Paul L. Boyer ◽  
Kevin Melody ◽  
Steven J. Smith ◽  
Linda L. Dunn ◽  
Chris Kline ◽  
...  
Keyword(s):  
Nucleic Acid ◽  
Reverse Transcriptase ◽  
Active Site ◽  
Nucleoside Analogs ◽  
Transcriptase Inhibitor ◽  
Drug Resistant ◽  
Resistance Mutations ◽  
Hydrophobic Region ◽  
Nnrti Resistance ◽  
Hiv 1

ABSTRACTTwo mutations, G112D and M230I, were selected in the reverse transcriptase (RT) of human immunodeficiency virus type 1 (HIV-1) by a novel nonnucleoside reverse transcriptase inhibitor (NNRTI). G112D is located near the HIV-1 polymerase active site; M230I is located near the hydrophobic region where NNRTIs bind. Thus, M230I could directly interfere with NNRTI binding but G112D could not. Biochemical and virological assays were performed to analyze the effects of these mutations individually and in combination. M230I alone caused a reduction in susceptibility to NNRTIs, while G112D alone did not. The G112D/M230I double mutant was less susceptible to NNRTIs than was M230I alone. In contrast, both mutations affected the ability of RT to incorporate nucleoside analogs. We suggest that the mutations interact with each other via the bound nucleic acid substrate; the nucleic acid forms part of the polymerase active site, which is near G112D. The positioning of the nucleic acid is influenced by its interactions with the “primer grip” region and could be influenced by the M230I mutation.IMPORTANCEAlthough antiretroviral therapy (ART) is highly successful, drug-resistant variants can arise that blunt the efficacy of ART. New inhibitors that are broadly effective against known drug-resistant variants are needed, although such compounds might select for novel resistance mutations that affect the sensitivity of the virus to other compounds. Compound 13 selects for resistance mutations that differ from traditional NNRTI resistance mutations. These mutations cause increased sensitivity to NRTIs, such as AZT.

scholarly journals A Motif Unique to the Human Dead-Box Protein DDX3 Is Important for Nucleic Acid Binding, ATP Hydrolysis, RNA/DNA Unwinding and HIV-1 Replication

PLoS ONE ◽  
2011 ◽  
Vol 6 (5) ◽  
pp. e19810 ◽  
Author(s):  
Anna Garbelli ◽  
Sandra Beermann ◽  
Giulia Di Cicco ◽  
Ursula Dietrich ◽  
Giovanni Maga
Keyword(s):  
Nucleic Acid ◽  
Atp Hydrolysis ◽  
Dna Unwinding ◽  
Nucleic Acid Binding ◽  
Dead Box ◽  
Hiv 1

scholarly journals Deaminase-independent inhibition of HIV-1 reverse transcription by APOBEC3G

2007 ◽  
Vol 35 (21) ◽  
pp. 7096-7108 ◽  
Author(s):  
Yasumasa Iwatani ◽  
Denise S.B. Chan ◽  
F. Wang ◽  
Kristen Stewart-Maynard ◽  
Wataru Sugiura ◽  
...  
Keyword(s):  
Nucleic Acid ◽  
Single Molecule ◽  
Reverse Transcription ◽  
Cytidine Deaminase ◽  
Rnase H ◽  
Host Protein ◽  
Nucleic Acid Binding ◽  
Nucleic Acid Chaperone ◽  
Kinetics Of ◽  
Hiv 1

Abstract APOBEC3G (A3G), a host protein that inhibits HIV-1 reverse transcription and replication in the absence of Vif, displays cytidine deaminase and single-stranded (ss) nucleic acid binding activities. HIV-1 nucleocapsid protein (NC) also binds nucleic acids and has a unique property, nucleic acid chaperone activity, which is crucial for efficient reverse transcription. Here we report the interplay between A3G, NC and reverse transcriptase (RT) and the effect of highly purified A3G on individual reactions that occur during reverse transcription. We find that A3G did not affect the kinetics of NC-mediated annealing reactions, nor did it inhibit RNase H cleavage. In sharp contrast, A3G significantly inhibited all RT-catalyzed DNA elongation reactions with or without NC. In the case of ( − ) strong-stop DNA synthesis, the inhibition was independent of A3G's catalytic activity. Fluorescence anisotropy and single molecule DNA stretching analyses indicated that NC has a higher nucleic acid binding affinity than A3G, but more importantly, displays faster association/disassociation kinetics. RT binds to ssDNA with a much lower affinity than either NC or A3G. These data support a novel mechanism for deaminase-independent inhibition of reverse transcription that is determined by critical differences in the nucleic acid binding properties of A3G, NC and RT.

scholarly journals Dynamics of Linker Residues Modulate the Nucleic Acid Binding Properties of the HIV-1 Nucleocapsid Protein Zinc Fingers

PLoS ONE ◽  
2014 ◽  
Vol 9 (7) ◽  
pp. e102150 ◽  
Author(s):  
Loussiné Zargarian ◽  
Carine Tisné ◽  
Pierre Barraud ◽  
Xiaoqian Xu ◽  
Nelly Morellet ◽  
...  
Keyword(s):  
Nucleic Acid ◽  
Nucleocapsid Protein ◽  
Zinc Fingers ◽  
Nucleic Acid Binding ◽  
Binding Properties ◽  
Hiv 1

scholarly journals Human Cellular Nucleic Acid-Binding Protein Zn2+ Fingers Support Replication of Human Immunodeficiency Virus Type 1 When They Are Substituted in the Nucleocapsid Protein

2003 ◽  
Vol 77 (15) ◽  
pp. 8524-8531 ◽  
Author(s):  
Connor F. McGrath ◽  
James S. Buckman ◽  
Tracy D. Gagliardi ◽  
William J. Bosche ◽  
Lori V. Coren ◽  
...  
Keyword(s):  
Human Immunodeficiency Virus ◽  
Nucleic Acid ◽  
Human Immunodeficiency Virus Type ◽  
Virus Type ◽  
Structural Characteristics ◽  
Nucleic Acid Binding ◽  
Genomic Rna ◽  
Immunodeficiency Virus ◽  
Hiv 1

ABSTRACT A family of cellular nucleic acid binding proteins (CNBPs) contains seven Zn2+ fingers that have many of the structural characteristics found in retroviral nucleocapsid (NC) Zn2+ fingers. The sequence of the NH2-terminal NC Zn2+ finger of the pNL4-3 clone of human immunodeficiency virus type 1 (HIV-1) was replaced individually with sequences from each of the seven fingers from human CNBP. Six of the mutants were normal with respect to protein composition and processing, full-length genomic RNA content, and infectivity. One of the mutants, containing the fifth CNBP Zn2+ finger (CNBP-5) packaged reduced levels of genomic RNA and was defective in infectivity. There appear to be defects in reverse transcription in the CNBP-5 infections. Models of Zn2+ fingers were constructed by using computational methods based on available structural data, and atom-atom interactions were determined by the hydropathic orthogonal dynamic analysis of the protein method. Defects in the CNBP-5 mutant could possibly be explained, in part, by restrictions of a set of required atom-atom interactions in the CNBP-5 Zn2+ finger compared to mutant and wild-type Zn2+ fingers in NC that support replication. The present study shows that six of seven of the Zn2+ fingers from the CNBP protein can be used as substitutes for the Zn2+ finger in the NH2-terminal position of HIV-1 NC. This has obvious implications in antiviral therapeutics and DNA vaccines employing NC Zn2+ finger mutants.

scholarly journals Two step binding of HIV-1 reverse transcriptase to nucleic acid substrates

1993 ◽  
Vol 21 (17) ◽  
pp. 3943-3949 ◽  
Author(s):  
Mogens KruhØfter ◽  
Claus Urbanke ◽  
Frank Grosse
Keyword(s):  
Nucleic Acid ◽  
Reverse Transcriptase ◽  
Hiv 1
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