Rational engineering enables co-crystallization and structural determination of the HIV-1 matrix-tRNA complex

Charles Bou-Nader; Jinwei Zhang

doi:10.1016/j.xpro.2021.101056

Rational engineering enables co-crystallization and structural determination of the HIV-1 matrix-tRNA complex

STAR Protoc. 2021 Dec 22;3(1):101056. doi: 10.1016/j.xpro.2021.101056. eCollection 2022 Mar 18.

Authors

Charles Bou-Nader¹, Jinwei Zhang¹

Affiliation

¹ Laboratory of Molecular Biology, National Institute of Diabetes and Digestive and Kidney Diseases, Bethesda, MD 20892, USA.

Abstract

Host tRNAs specifically interact with the matrix domain (MA) of HIV-1 major structural polyprotein, Gag, to control its membrane localization and virion assembly. In this protocol, we describe the purification and engineering of HIV-1 MA and tRNA, and the co-crystallization and structure determination of the complex using X-ray crystallography. Rational engineering of the tRNA surface created tRNA-tRNA packing contacts that drove the formation of diffraction-quality co-crystals. This protocol can be adapted to solve other ribonucleoprotein complex structures containing structured RNAs. For complete details on the use and execution of this protocol, please refer to Bou-Nader et al. (2021).

Keywords: Biophysics; Microbiology; Molecular Biology; Structural Biology; X-ray Crystallography.

Publication types

Research Support, N.I.H., Intramural
Research Support, U.S. Gov't, Non-P.H.S.

MeSH terms

Crystallization
HIV-1* / genetics
RNA, Transfer / genetics
Virus Assembly
gag Gene Products, Human Immunodeficiency Virus / genetics

Substances

gag Gene Products, Human Immunodeficiency Virus
RNA, Transfer

Grants and funding

ZIA DK075136/ImNIH/Intramural NIH HHS/United States