A lysine ring in HIV capsid pores coordinates IP6 to drive mature capsid assembly.

Change log
Renner, Nadine 
Mallery, Donna L 
Faysal, KM Rifat 
Peng, Wang 

The HIV capsid self-assembles a protective conical shell that simultaneously prevents host sensing whilst permitting the import of nucleotides to drive DNA synthesis. This is accomplished through the construction of dynamic, highly charged pores at the centre of each capsid multimer. The clustering of charges required for dNTP import is strongly destabilising and it is proposed that HIV uses the metabolite IP6 to coordinate the pore during assembly. Here we have investigated the role of inositol phosphates in coordinating a ring of positively charged lysine residues (K25) that forms at the base of the capsid pore. We show that whilst IP5, which can functionally replace IP6, engages an arginine ring (R18) at the top of the pore, the lysine ring simultaneously binds a second IP5 molecule. Dose dependent removal of K25 from the pore severely inhibits HIV infection and concomitantly prevents DNA synthesis. Cryo-tomography reveals that K25A virions have a severe assembly defect that inhibits the formation of mature capsid cones. Monitoring both the kinetics and morphology of capsids assembled in vitro reveals that while mutation K25A can still form tubes, the ability of IP6 to drive assembly of capsid cones has been lost. Finally, in single molecule TIRF microscopy experiments, capsid lattices in permeabilised K25 mutant virions are rapidly lost and cannot be stabilised by IP6. These results suggest that the coordination of IP6 by a second charged ring in mature hexamers drives the assembly of conical capsids capable of reverse transcription and infection.

Capsid, Cell Line, DNA, Viral, HIV-1, Humans, Lysine, Microscopy, Fluorescence, Nucleotides, Phytic Acid, Virus Assembly
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PLoS Pathog
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Public Library of Science (PLoS)
Medical Research Council (UK) (U105181010)
Wellcome Trust (200594/Z/16/Z)
Wellcome Trust (200594/Z/16/Z)
Wellcome Trust (214344/A/18/Z)
Wellcome Trust (214344/A/18/Z)
National Health and Medical Research Council (APP1100771)
National Health and Medical Research Council (APP1100771)
InstitNational Health and Medical Research Council (APP1100771)
National Health and Medical Research Council (APP1158338)
National Health and Medical Research Council (APP1158338)
Australian Research Council (ARC LIEF 190100165)