학술논문

Investigation of HIV-1 Gag binding with RNAs and lipids using Atomic Force Microscopy.
Document Type
article
Source
PloS one. 15(2)
Subject
Cell Membrane
Humans
HIV-1
HIV Infections
HIV Seropositivity
Multiprotein Complexes
Lipids
Phosphatidylinositol 4
5-Diphosphate
RNA-Binding Proteins
RNA
Viral
Microscopy
Atomic Force
Protein Binding
gag Gene Products
Human Immunodeficiency Virus
Protein Multimerization
q-bio.BM
Phosphatidylinositol 4
5-Diphosphate
RNA
Viral
Microscopy
Atomic Force
gag Gene Products
Human Immunodeficiency Virus
General Science & Technology
Language
Abstract
Atomic Force Microscopy was utilized to study the morphology of Gag, ΨRNA, and their binding complexes with lipids in a solution environment with 0.1Å vertical and 1nm lateral resolution. TARpolyA RNA was used as a RNA control. The lipid used was phospha-tidylinositol-(4,5)-bisphosphate (PI(4,5)P2). The morphology of specific complexes Gag-ΨRNA, Gag-TARpolyA RNA, Gag-PI(4,5)P2 and PI(4,5)P2-ΨRNA-Gag were studied. They were imaged on either positively or negatively charged mica substrates depending on the net charges carried. Gag and its complexes consist of monomers, dimers and tetramers, which was confirmed by gel electrophoresis. The addition of specific ΨRNA to Gag is found to increase Gag multimerization. Non-specific TARpolyA RNA was found not to lead to an increase in Gag multimerization. The addition PI(4,5)P2 to Gag increases Gag multimerization, but to a lesser extent than ΨRNA. When both ΨRNA and PI(4,5)P2 are present Gag undergoes comformational changes and an even higher degree of multimerization.