Genome-wide CRISPR knockout screen identifies ZNF304 as a silencer of HIV transcription that promotes viral latency

PLoS Pathog. 2020 Sep 21;16(9):e1008834. doi: 10.1371/journal.ppat.1008834. eCollection 2020 Sep.

Abstract

Despite the widespread use of anti-retroviral therapy, human immunodeficiency virus (HIV) still persists in an infected cell reservoir that harbors transcriptionally silent yet replication-competent proviruses. While significant progress has been made in understanding how the HIV reservoir is established, transcription repression mechanisms that are enforced on the integrated viral promoter have not been fully revealed. In this study, we performed a whole-genome CRISPR knockout screen in HIV infected T cells to identify host genes that potentially promote HIV latency. Of several top candidates, the KRAB-containing zinc finger protein, ZNF304, was identified as the top hit. ZNF304 silences HIV gene transcription through associating with TRIM28 and recruiting to the viral promoter heterochromatin-inducing methyltransferases, including the polycomb repression complex (PRC) and SETB1. Depletion of ZNF304 expression reduced levels of H3K9me3, H3K27me3 and H2AK119ub repressive histone marks on the HIV promoter as well as SETB1 and TRIM28, ultimately enhancing HIV gene transcription. Significantly, ZNF304 also promoted HIV latency, as its depletion delayed the entry of HIV infected cells into latency. In primary CD4+ cells, ectopic expression of ZNF304 silenced viral transcription. We conclude that by associating with TRIM28 and recruiting host transcriptional repressive complexes, SETB1 and PRC, to the HIV promoter, ZNF304 silences HIV gene transcription and promotes viral latency.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • CD4-Positive T-Lymphocytes / metabolism*
  • CD4-Positive T-Lymphocytes / virology
  • CRISPR-Cas Systems
  • Gene Expression Regulation, Viral*
  • Gene Knockout Techniques
  • Gene Silencing*
  • Genome-Wide Association Study
  • HIV-1 / physiology*
  • Humans
  • Jurkat Cells
  • Promoter Regions, Genetic
  • Repressor Proteins* / genetics
  • Repressor Proteins* / metabolism
  • Transcription Factors* / genetics
  • Transcription Factors* / metabolism
  • Transcription, Genetic*
  • Tripartite Motif-Containing Protein 28 / genetics
  • Tripartite Motif-Containing Protein 28 / metabolism
  • Virus Latency*

Substances

  • Repressor Proteins
  • Transcription Factors
  • ZNF304 protein, human
  • TRIM28 protein, human
  • Tripartite Motif-Containing Protein 28

Grants and funding

This work was funded by the Israel Science Foundation (ISF; https://www.isf.org.il/#/) for RT (755/17). The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.