The p66 immature precursor of HIV-1 reverse transcriptase.
In contrast to the wealth of structural data available for the mature p66/p51 heterodimeric human immunodeficiency virus type 1 reverse transcriptase (RT), the structure of the homodimeric p66 precursor remains unknown. In all X-ray structures of mature RT, free or complexed, the processing site in the p66 subunit, for generating the p51 subunit, is sequestered into a β-strand within the folded ribonuclease H (RNH) domain and is not readily accessible to proteolysis, rendering it difficult to propose a simple and straightforward mechanism of the maturation step. Here, we investigated, by solution NMR, the conformation of the RT p66 homodimer. Our data demonstrate that the RNH and Thumb domains in the p66 homodimer are folded and possess conformations very similar to those in mature RT. This finding suggests that maturation models which invoke a complete or predominantly unfolded RNH domain are unlikely. The present study lays the foundation for further in-depth mechanistic investigations at the atomic level.
Department of Structural Biology, University of Pittsburgh School of Medicine, Pittsburgh, Pennsylvania, 15260.
Naima G
NG
Sharaf
Eric
E
Poliner
Ryan L
RL
Slack
Martin T
MT
Christen
In-Ja L
IJ
Byeon
Michael A
MA
Parniak
Angela M
AM
Gronenborn
Rieko
R
Ishima
D001665:Binding Sites; D054303:HIV Reverse Transcriptase / Q000737:chemistry*; D006801:Humans; D008958:Models, Molecular; D011487:Protein Conformation; D055503:Protein Multimerization; D011498:Protein Precursors / Q000737:chemistry*; D016914:Ribonuclease H / Q000737:chemistry*
D011498:Protein Precursors; C514824:reverse transcriptase, Human immunodeficiency virus 1; D054303:HIV Reverse Transcriptase; D016914:Ribonuclease H
NMR; dynamics; enzyme; maturation; protein; proteolysis; structure