Protocol for an in vitro assay to study HIV-1 Tat methylation

Abstract

Summary: A critical virus-encoded regulator of HIV-1 transcription is the Tat protein, which is required to potently activate transcription. Tat is regulated by a wide variety of post-translational modifications. This protocol describes an in vitro assay to study Tat methylation. We describe steps for incorporation of radioactive methyl groups into Tat protein, visualization by gel analysis, Coomassie blue stain, gel drying, and detection by autoradiography. This protocol can also be used to assess methylation in other proteins such as histones.For complete details on the use and execution of this protocol, please refer to Boehm et al. (2023).1 : Publisher’s note: Undertaking any experimental protocol requires adherence to local institutional guidelines for laboratory safety and ethics.

Keywords

• Cell Biology
• Molecular Biology
• Protein Biochemistry