Annealing of ssDNA and compaction of dsDNA by the HIV-1 nucleocapsid and Gag proteins visualized using nanofluidic channels
Artikel i vetenskaplig tidskrift, 2019

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.

nucleocapsid

nanofluidics

single DNA molecules

Gag

HIV-1

Författare

Kai Jiang

Chalmers, Biologi och bioteknik, Livsmedelsvetenskap

Sriram Kesarimangalam

Chalmers, Biologi och bioteknik, Kemisk biologi

Yii Lih Lin

Chalmers, Biologi och bioteknik, Kemisk biologi

Fredrik Westerlund

Chalmers, Biologi och bioteknik, Kemisk biologi

Quarterly Reviews of Biophysics

0033-5835 (ISSN) 1469-8994 (eISSN)

Vol. 52

Ämneskategorier

Biofysik

Nanoteknik

DOI

10.1017/S0033583518000124

Mer information

Skapat

2019-05-28