Single-base m⁶A epitranscriptomics reveals novel HIV-1 host interaction targets in primary CD4⁺ T cells

Area of Science:

• * Molecular Biology
• * Virology
• * Immunology

Background:

• * N-methyladenosine (m6A) is a key RNA modification regulating gene expression.
• * m6A modification influences viral and cellular gene expression during HIV-1 infection.
• * The role of m6A in HIV-1 infection of primary CD4+ T cells is not fully understood.

Purpose of the Study:

• * To investigate the function of m6A modification in HIV-1 infection of primary CD4+ T cells.
• * To identify cellular and viral genes affected by m6A modification during HIV-1 infection.
• * To elucidate the role of perilipin 3 (PLIN3) in HIV-1 replication.

Main Methods:

• * Single-base m6A-specific RNA sequencing to identify m6A-modified mRNAs.
• * Analysis of m6A sites in HIV-1 RNA.
• * Measurement of PLIN3 mRNA and protein levels, nuclear accumulation, and translation efficiency.
• * PLIN3 knockdown experiments in primary CD4+ T cells.

Main Results:

• * HIV-1 infection promotes interaction between METTL3/METTL14 m6A writer complex subunits.
• * Identified differentially m6A-modified cellular mRNAs, including PLIN3, and 30 m6A sites in HIV-1 RNA.
• * HIV-1 infection increased PLIN3 mRNA levels and nuclear accumulation but decreased PLIN3 protein expression and translation.
• * PLIN3 knockdown reduced HIV-1 release but enhanced virion infectivity.

Conclusions:

• * m6A RNA modification is important for regulating HIV-1 infection in primary CD4+ T cells.
• * PLIN3 is identified as a regulatory protein in HIV-1 replication.
• * Findings suggest a novel regulatory mechanism involving m6A modification and PLIN3 in HIV-1 pathogenesis.