vaccinia virus–encoded ribonucleotide reductase subunits are differentially required for replication and pathogenesis牛痘病毒编码核苷酸还原酶亚基不同需要复制和发病机理.pdfVIP

Vaccinia Virus–Encoded Ribonucleotide Reductase Subunits Are Differentially Required for Replication and Pathogenesis 1 1 2 2 3 Don B. Gammon , Branawan Gowrishankar , Sophie Duraffour , Graciela Andrei , Chris Upton , David H. Evans1* 1 Department of Medical Microbiology and Immunology, University of Alberta, Edmonton, Alberta, Canada, 2 Laboratory of Virology and Rega Institute for Medical Research, Katholieke Universiteit Leuven, Leuven, Belgium, 3 Biochemistry and Microbiology, University of Victoria, Victoria, British Columbia, Canada Abstract Ribonucleotide reductases (RRs) are evolutionarily-conserved enzymes that catalyze the rate-limiting step during dNTP synthesis in mammals. RR consists of both large (R1) and small (R2) subunits, which are both required for catalysis by the R12R22 heterotetrameric complex. Poxviruses also encode RR proteins, but while the Orthopoxviruses infecting humans [e.g. vaccinia (VACV), variola, cowpox, and monkeypox viruses] encode both R1 and R2 subunits, the vast majority of Chordopoxviruses encode only R2 subunits. Using plaque morphology, growth curve, and mouse model studies, we investigated the requirement of VACV R1 (I4) and R2 (F4) subunits for replication and pathogenesis using a panel of mutant viruses in which one or more viral RR genes had been inactivated. Surprisingly, VACV F4, but not I4, was required for efficient replication in culture and virulence in mice. The growth defects of VACV strains lacking F4 could be complemented by genes encoding other Chordopoxvirus R2 subunits, suggesting conservation of function between poxvirus R2 proteins. Expression of F4 proteins encoding a point mutation pr