Insights into the evolutionary history of the virulent factor HBHA of Mycobacterium tuberculosis

In Mycobacterium tuberculosis, heparin-binding hemagglutinin (HBHAMT) has a relevant role in infection. It is also present in non-virulent mycobacteria and ancient actinobacteria, such as Rhodococcus opacus. To have a better understanding of the underlying mechanisms that shaped the evolutionary divergence of these proteins, we performed a comprehensive phylogenetic analysis of the regulatory sequences that drive the expression of hbha in saprophytic and pathogenic mycobacterial species. The alignment of the hbha loci showed the appearance of intergenic sequences containing regulatory elements upstream the hbha gene; this sequence arrangement is present only in slow-growing pathogenic mycobacteria. The heterologous expression of HBHAMT in oleaginous R. opacus PD630 results in protein binding to lipid droplets, as it happens with HBHA proteins from saprophytic mycobacteria. We hypothesize that mycobacterial hbha gene cluster underwent functional divergence during the evolutionary differentiation of slow-growing pathogenic mycobacteria. We propose here an evolutionary scenario to explain the structural and functional divergence of HBHA in fast and slow-growing mycobacteria.