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Oligomeric structures of poliovirus polymerase are important for function
Author(s) -
Hobson Scott D.,
Rosenblum Eric S.,
Richards Oliver C.,
Richmond Kathryn,
Kirkegaard Karla,
Schultz Steve C.
Publication year - 2001
Publication title -
the embo journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 7.484
H-Index - 392
eISSN - 1460-2075
pISSN - 0261-4189
DOI - 10.1093/emboj/20.5.1153
Subject(s) - polymerase , biology , rna polymerase , rna dependent rna polymerase , rna polymerase i , rna polymerase ii , rna polymerase iii , poliovirus , microbiology and biotechnology , rna , virology , biochemistry , enzyme , virus , gene expression , gene , promoter
Central to the replication of poliovirus and other positive‐strand RNA viruses is the virally encoded RNA‐dependent RNA polymerase. Previous biochemical studies have suggested that direct polymerase–polymerase interactions might be important for polymerase function, and the structure of poliovirus polymerase has revealed two regions of extensive polymerase–polymerase interaction. To explore potential functional roles for the structurally observed polymerase–polymerase interactions, we have performed RNA binding and extension studies of mutant polymerase proteins in solution, disulfide cross‐linking studies, mutational analyses in cells, in vitro activity analyses and RNA substrate modeling studies. The results of these studies indicate that both regions of polymerase–polymerase interaction observed in the crystals are indeed functionally important and, furthermore, reveal specific functional roles for each. One of the two regions of interaction provides for efficient substrate RNA binding and the second is crucial for forming catalytic sites. These studies strongly support the hypothesis that the polymerase–polymerase interactions discovered in the crystal structure provide an exquisitely detailed structural context for poliovirus polymerase function and for poliovirus RNA replication in cells.