3KLF

Crystal structure of wild-type HIV-1 Reverse Transcriptase crosslinked to a DSDNA with a bound excision product, AZTPPPPA

Summary for 3KLF

• Entry DOI: 10.2210/pdb3klf/pdb
• Related: 3KLE 3KLG 3KLH 3KLI
• Descriptor: Reverse transcriptase/ribonuclease H, p51 RT, DNA (5'-D(*AP*T*GP*CP*AP*TP*GP*GP*CP*GP*CP*CP*CP*GP*AP*AP*CP*AP*GP*GP*GP*AP*CP*TP*GP*TP*G)-3'), ... (8 entities in total)
• Functional Keywords: azt resistance mechanism, hiv-1 reverse transcriptase, wild-type, azt resistance mutations, p51/p66, nuceleoside inhibitor, thymidine analog mutations, aids, hiv, dna polymerase, nrti, nrti resistance, azt, aztppppa, aztp4a, dinucleoside tetraphosphate, dna-directed dna polymerase, endonuclease, hydrolase, magnesium, metal-binding, multifunctional enzyme, nuclease, rna-directed dna polymerase, transferase, transferase-dna complex, transferase/dna
• Biological source: Human immunodeficiency virus type 1 (HIV-1); More
• Cellular location: Matrix protein p17: Virion (Potential). Capsid protein p24: Virion (Potential). Nucleocapsid protein p7: Virion (Potential). Reverse transcriptase/ribonuclease H: Virion (Potential). Integrase: Virion (Potential): P03366 P03366
• Total number of polymer chains: 16
• Total formula weight: 527063.14

Authors

Tu, X.,Das, K.,Sarafianos, S.G.,Arnold, E. (deposition date: 2009-11-07, release date: 2010-09-22, Last modification date: 2024-10-09)

Primary citation

Tu, X.,Das, K.,Han, Q.,Bauman, J.D.,Clark, A.D.,Hou, X.,Frenkel, Y.V.,Gaffney, B.L.,Jones, R.A.,Boyer, P.L.,Hughes, S.H.,Sarafianos, S.G.,Arnold, E.
Structural basis of HIV-1 resistance to AZT by excision.
Nat.Struct.Mol.Biol., 17:1202-1209, 2010

PubMed Abstract: Human immunodeficiency virus (HIV-1) develops resistance to 3'-azido-2',3'-deoxythymidine (AZT, zidovudine) by acquiring mutations in reverse transcriptase that enhance the ATP-mediated excision of AZT monophosphate from the 3' end of the primer. The excision reaction occurs at the dNTP-binding site, uses ATP as a pyrophosphate donor, unblocks the primer terminus and allows reverse transcriptase to continue viral DNA synthesis. The excision product is AZT adenosine dinucleoside tetraphosphate (AZTppppA). We determined five crystal structures: wild-type reverse transcriptase-double-stranded DNA (RT-dsDNA)-AZTppppA; AZT-resistant (AZTr; M41L D67N K70R T215Y K219Q) RT-dsDNA-AZTppppA; AZTr RT-dsDNA terminated with AZT at dNTP- and primer-binding sites; and AZTr apo reverse transcriptase. The AMP part of AZTppppA bound differently to wild-type and AZTr reverse transcriptases, whereas the AZT triphosphate part bound the two enzymes similarly. Thus, the resistance mutations create a high-affinity ATP-binding site. The structure of the site provides an opportunity to design inhibitors of AZT-monophosphate excision.

PubMed: 20852643
DOI: 10.1038/nsmb.1908

Experimental method

X-RAY DIFFRACTION (3.15 Å)