Research

Protein methylation is required to maintain optimal HIV-1 infectivity

Nicole M Willemsen^1†, Eleanor M Hitchen^2†, Tracey J Bodetti¹, Ann Apolloni¹, David Warrilow¹, Sabine C Piller² and David Harrich^1*

• * Corresponding author: David Harrich davidH@qimr.edu.au

• † Equal contributors

Author Affiliations

¹ Division of Immunology and Infectious Disease, Queensland Institute of Medical Research, Brisbane, Queensland, 4006, Australia

² HIV Protein Functions and Interactions Group, Centre for Virus Research, Westmead Millennium Institute, Westmead NSW 2145, Australia

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Retrovirology 2006, 3:92 doi:10.1186/1742-4690-3-92

Published: 15 December 2006

Abstract

Background:

Protein methylation is recognized as a major protein modification pathway regulating diverse cellular events such as protein trafficking, transcription, and signal transduction. More recently, protein arginine methyltransferase activity has been shown to regulate HIV-1 transcription via Tat. In this study, adenosine periodate (AdOx) was used to globally inhibit protein methyltransferase activity so that the effect of protein methylation on HIV-1 infectivity could be assessed.

Results:

Two cell culture models were used: HIV-1-infected CEM T-cells and HEK293T cells transfected with a proviral DNA plasmid. In both models, AdOx treatment of cells increased the levels of virion in culture supernatant. However, these viruses had increased levels of unprocessed or partially processed Gag-Pol, significantly increased diameter, and displayed reduced infectivity in a MAGI X4 assay. AdOx reduced infectivity equally in both dividing and non-dividing cells. However, infectivity was further reduced if Vpr was deleted suggesting virion proteins, other than Vpr, were affected by protein methylation. Endogenous reverse transcription was not inhibited in AdOx-treated HIV-1, and infectivity could be restored by pseudotyping HIV with VSV-G envelope protein. These experiments suggest that AdOx affects an early event between receptor binding and uncoating, but not reverse transcription.

Conclusion:

Overall, we have shown for the first time that protein methylation contributes towards maximal virus infectivity. Furthermore, our results also indicate that protein methylation regulates HIV-1 infectivity in a complex manner most likely involving the methylation of multiple viral or cellular proteins and/or multiple steps of replication.