HIV-1 Nucleocapsid Protein Binds Double-Stranded DNA in Multiple Modes to Regulate Compaction and Capsid Uncoating

Viruses. 2022 Jan 25;14(2):235. doi: 10.3390/v14020235.

Abstract

The HIV-1 nucleocapsid protein (NC) is a multi-functional protein necessary for viral replication. Recent studies have demonstrated reverse transcription occurs inside the fully intact viral capsid and that the timing of reverse transcription and uncoating are correlated. How a nearly 10 kbp viral DNA genome is stably contained within a narrow capsid with diameter similar to the persistence length of double-stranded (ds) DNA, and the role of NC in this process, are not well understood. In this study, we use optical tweezers, fluorescence imaging, and atomic force microscopy to observe NC binding a single long DNA substrate in multiple modes. We find that NC binds and saturates the DNA substrate in a non-specific binding mode that triggers uniform DNA self-attraction, condensing the DNA into a tight globule at a constant force up to 10 pN. When NC is removed from solution, the globule dissipates over time, but specifically-bound NC maintains long-range DNA looping that is less compact but highly stable. Both binding modes are additionally observed using AFM imaging. These results suggest multiple binding modes of NC compact DNA into a conformation compatible with reverse transcription, regulating the genomic pressure on the capsid and preventing premature uncoating.

Keywords: DNA condensation; HIV-1 nucleocapsid protein; atomic force microscopy; capsid uncoating; optical tweezers.

Publication types

  • Research Support, N.I.H., Extramural
  • Research Support, U.S. Gov't, Non-P.H.S.

MeSH terms

  • DNA / chemistry
  • DNA / metabolism*
  • HIV-1 / genetics
  • HIV-1 / metabolism
  • HIV-1 / physiology*
  • Microscopy, Atomic Force
  • Nucleic Acid Conformation
  • Nucleocapsid Proteins / metabolism*
  • Protein Binding
  • Reverse Transcription
  • Virus Replication
  • Virus Uncoating*
  • gag Gene Products, Human Immunodeficiency Virus / metabolism*

Substances

  • NCP7 protein, Human immunodeficiency virus 1
  • Nucleocapsid Proteins
  • gag Gene Products, Human Immunodeficiency Virus
  • DNA