Functional Characterization of EngAMS, a P-Loop GTPase of Mycobacterium smegmatisReportar como inadecuado

Functional Characterization of EngAMS, a P-Loop GTPase of Mycobacterium smegmatis - Descarga este documento en PDF. Documentación en PDF para descargar gratis. Disponible también para leer online.

Bacterial P-loop GTPases belong to a family of proteins that selectively hydrolyze a small molecule guanosine tri-phosphate GTP to guanosine di-phosphate GDP and inorganic phosphate, and regulate several essential cellular activities such as cell division, chromosomal segregation and ribosomal assembly. A comparative genome sequence analysis of different mycobacterial species indicates the presence of multiple P-loop GTPases that exhibit highly conserved motifs. However, an exact function of most of these GTPases in mycobacteria remains elusive. In the present study we characterized the function of a P-loop GTPase in mycobacteria by employing an EngA homologue from Mycobacterium smegmatis, encoded by an open reading frame, designated as MSMEG 3738. Amino acid sequence alignment and phylogenetic analysis suggest that MSMEG 3738 termed as EngAMS is highly conserved in mycobacteria. Homology modeling of EngAMS reveals a cloverleaf structure comprising of α-β fold typical to EngA family of GTPases. Recombinant EngAMS purified from E. coli exhibits a GTP hydrolysis activity which is inhibited by the presence of GDP. Interestingly, the EngAMS protein is co-eluted with 16S and 23S ribosomal RNA during purification and exhibits association with 30S, 50S and 70S ribosomal subunits. Further studies demonstrate that GTP is essential for interaction of EngAMS with 50S subunit of ribosome and specifically C-terminal domains of EngAMS are required to facilitate this interaction. Moreover, EngAMS devoid of N-terminal region interacts well with 50S even in the absence of GTP, indicating a regulatory role of the N-terminal domain in EngAMS-50S interaction.

Autor: Nisheeth Agarwal , Madhu Pareek, Preeti Thakur, Vibha Pathak



Documentos relacionados