Molecular Mechanisms behind Conformational Transitions of Influenza Virus Hemagglutinin Membrane Anchor

Membrane fusion is a fundamental process exploited by enveloped viruses to enter host cells. In the case of influenza virus, fusion is facilitated by the trimeric viral hemagglutinin protein (HA). So far, major focus has been put on its N-terminal fusion peptides, which are directly responsible for fusion initiation. A growing body of evidence points also to a significant functional role of HA C-terminal domain, which however remains incompletely understood. Our computational study aimed to elucidate the structural and functional interdependencies within the HA C-terminal region, encompassing the transmembrane domain (TMD) and the cytoplasmic tail (CT). In particular, we were interested in conformational shift of the TMD in response to varying cholesterol concentration in the viral membrane and in its modulation by the presence of CT. Using free energy calculations based on atomistic molecular dynamics simulations, we characterized transitions between straight and tilted metastable TMD configurations under varying conditions. We found that the presence of CT is essential for achieving a stable, highly tilted TMD configuration. As we demonstrate, such configuration of HA membrane anchor likely supports the tilting motion of its ectodomain which needs to be executed during membrane fusion. This finding highlights the functional role of, so far relatively overlooked, CT region.