The Cys3 -His1 Zinc binding motif of the Respiratory Syncytial Virus M2-1 tetramer modulates its dissociation to folded APO-monomers

ESPERANTE SEBASTIÁN A.; NOVAL MARÍA GABRIELA; ANTONELLA TAMARA; DE PRAT-GAY GONZALO

Granada

Congreso; 15th International Negative Strand Virus Meeting; 2013

Human Respiratory Syncytial Virus (hRSV)is a major cause of pediatric viral respiratory tract infections and is the leading cause of infant hospitalization in industrialized countries. The RSV virus belongs to the order Mononegavirales, family Paramyxoviridae,genusPneumovirus. The RSV genome consists of a ~15 kb single stranded non segmented nega2ve-­‐sense RNA which is encapsidated by the nucleocapsid (N)protein. The resulting ribonucleoprotein complex(N-­‐RNA)is the template for transcription and replication of the viral genome by theRNA-­‐dependent RNA polymerase complex which comprises the large polymerase protein L,the phosphoprotein P and the transcription antiterminator factor M2-­‐1. The hRSV M2-­‐1 is a tight tetramer bearing a Cys3-­‐His1 motif which binds one zinc atom per monomer. The CCCH motif is found in the M2-­‐1 proteins of all Pneumoviruses and is essential to maintain the functional integrity of M2-­‐1 and for viral viability. However, the exact role that plays the CCCH motif in M2-­‐1 protein structure and function is still unknown. In the present study, we expressed and purified the hRSV M2-­‐1 protein in bacteria and investigated the role of Zinc in its conformational properties and stability in solution.