Mycobacterium tuberculosis which is the causative agent of the number-one killer disease worldwide. In addition to Mtb, the genus Mycobacterium includes some of the well-known Actinobacteria, such as the well-studied model organism Mycobacterium smegmatis, Mycobacterium abscessus, Mycobacterium leprae, and a large number of Streptomyces species. In recent years, studies on transcription initiation, elongation and termination have been carried out in Mtb and other members. These studies, though not as exhaustive as in E. coli, have revealed considerable differences from the E. coli paradigm. For example, the Mtb and other mycobacterial genomes code for a larger number of sigma factors as compared to E.coli and also for the several transcription factors unique to mycobacteria. Several promoters and the mechanism of gene expression regulation have been studied. Absence of ATrich UP elements and GC-rich sequences in discriminator sequences in the promoters contribute to the differences in promoter-polymerase interaction and its regulation. Additionally, attempts have been made to elucidate features of RNAP from Mycobacterium SAR131675 species and the transcription elongation rates also appear to vary between different RNAPs. Furthermore, the scarcity of canonical intrinsic terminators and an abundance of noncanonical intrinsic terminators across mycobacteria also suggest differences in the transcription termination machinery. Given the dissimilarities in various steps of transcription between mycobacteria and E. coli, it is likely that mycobacterial Rho homologs have also evolved to function differently and optimally for their specific cellular context. Notably, sequence analysis showed that the Rho homologs in Mycobacterium species and other actinobacteria are larger than EcRho mainly due to an ‘extra-stretch’ of,150–200 residues in their RNA-binding domains. In this manuscript, we present results demonstrating that purified M. tuberculosis Rho can hydrolyse purine nucleoside triphosphates – ATP and GTPin presence of mycobacterial RNA. The extended N-terminal region of MtbRho, having a distinct RNA-binding ‘subdomain’, can itself interact with RNA and may contribute to the overall interaction. The MtbRho-RNA interactions are stable and the interactions induce changes in the conformation and oligomerization status of the protein. Notably, the Rho homologs from actinobacteria form a distinct branch and the Nterminal halves of actinobacterial rho proteins contain an ‘extrastretch/subdomain’ of 150–200 residues. The sequence composition of this fragment includes a large number of Arg, Asp, Asn and Gly residues but very few hydrophobic and aromatic amino acids. Although no function can be conclusively predicted from the sequence analysis, its role in interacting with RNA is postulated from the presence of a large number of basic amino acid residues.