Mutations in DNA Binding and Transactivation Domains Affect the Dynamics of Parvovirus NS1 Protein
| Autor: | Olli Kalliolinna, Einari A. Niskanen, Maija Vihinen-Ranta, Teemu O. Ihalainen, Milla M. Häkkinen |
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| Jazyk: | angličtina |
| Rok vydání: | 2013 |
| Předmět: |
HMG-box
Parvovirus Canine viruses Immunology DNA Mutational Analysis Mutation Missense NS1 proteiini Viral Nonstructural Proteins Virus Replication Microbiology NS1 protein Single-stranded binding protein Cell Line SeqA protein domain Virology Animals DNA binding Replication protein A biology Ter protein parvovirus virus diseases DNA n sitoutuminen [DNA] biochemical phenomena metabolism and nutrition Molecular biology Cell biology Virus-Cell Interactions Protein Structure Tertiary DNA binding site DNA-Binding Proteins Insect Science biology.protein Mutant Proteins Viral genome replication Binding domain Protein Binding |
| Popis: | The multifunctional replication protein of autonomous parvoviruses, NS1, is vital for viral genome replication and for the control of viral protein production. Two DNA-interacting domains of NS1, the N-terminal and helicase domains, are necessary for these functions. In addition, the N and C termini of NS1 are required for activation of viral promoter P38. By comparison with the structural and biochemical data from other parvoviruses, we identified potential DNA-interacting amino acid residues from canine parvovirus NS1. The role of the identified amino acids in NS1 binding dynamics was studied by mutagenesis, fluorescence recovery after photobleaching, and computer simulations. Mutations in the predicted DNA-interacting amino acids of the N-terminal and helicase domains increased the intranuclear binding dynamics of NS1 dramatically. A substantial increase in binding dynamics was also observed for NS1 mutants that targeted the metal ion coordination site in the N terminus. Interestingly, contrary to other mutants, deletion of the C terminus resulted in slower binding dynamics of NS1. P38 transactivation was severely reduced in both N-terminal DNA recognition and in C-terminal deletion mutants. These data suggest that the intranuclear dynamics of NS1 are largely characterized by its sequence-specific and -nonspecific binding to double-stranded DNA. Moreover, binding of NS1 is equally dependent on the N-terminal domain and conserved β-loop of the helicase domain. |
| Databáze: | OpenAIRE |
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