Annealing of ssDNA and compaction of dsDNA by the HIV-1 nucleocapsid and Gag proteins visualized using nanofluidic channels - CNRS - Centre national de la recherche scientifique Accéder directement au contenu
Article Dans Une Revue Quarterly Reviews of Biophysics Année : 2019

Annealing of ssDNA and compaction of dsDNA by the HIV-1 nucleocapsid and Gag proteins visualized using nanofluidic channels

Kai Jiang
  • Fonction : Auteur
Sriram Kk
Thiebault Lequeu
  • Fonction : Auteur
Yii-Lih Lin
  • Fonction : Auteur
Fredrik Westerlund

Résumé

The nucleocapsid protein NC is a crucial component in the human immunodeficiency virus type 1 life cycle. It functions both in its processed mature form and as part of the polyprotein Gag that plays a key role in the formation of new viruses. NC can protect nucleic acids (NAs) from degradation by compacting them to a dense coil. Moreover, through its NA chaperone activity, NC can also promote the most stable conformation of NAs. Here, we explore the balance between these activities for NC and Gag by confining DNA-protein complexes in nanochannels. The chaperone activity is visualized as concatemerization and circularization of long DNA via annealing of short single-stranded DNA overhangs. The first ten amino acids of NC are important for the chaperone activity that is almost completely absent for Gag. Gag condenses DNA more efficiently than mature NC, suggesting that additional residues of Gag are involved. Importantly, this is the first single DNA molecule study of full-length Gag and we reveal important differences to the truncated Δ-p6 Gag that has been used before. In addition, the study also highlights how nanochannels can be used to study reactions on ends of long single DNA molecules, which is not trivial with competing single DNA molecule techniques.

Domaines

Biotechnologies

Dates et versions

hal-03636055 , version 1 (08-04-2022)

Identifiants

Citer

Kai Jiang, Nicolas Humbert, Sriram Kk, Thiebault Lequeu, Yii-Lih Lin, et al.. Annealing of ssDNA and compaction of dsDNA by the HIV-1 nucleocapsid and Gag proteins visualized using nanofluidic channels. Quarterly Reviews of Biophysics, 2019, 52, ⟨10.1017/S0033583518000124⟩. ⟨hal-03636055⟩

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