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

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).

SUBMITTER: Bou-Nader C

PROVIDER: S-EPMC8715211 | biostudies-literature | 2022 Mar

REPOSITORIES: biostudies-literature

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

Bou-Nader Charles C Zhang Jinwei J

STAR protocols 20211222 1

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 ca ...[more]

PMID: 35005638

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