Non-covalent interaction between nucleocapsid protein of Tula hantavirus and small ubiquitin-related modifier-1, SUMO-1.
To find cellular binding counterparts for the nucleocapsid protein (N) of Tula hantavirus (TULV), two cDNA libraries were screened using yeast two-hybrid systems based on LexA and Gal4 transcription factors. Five cDNA clones encoding SUMO-1 (Small Ubiquitin-related MOdifier, also known as sentrin) were selected in the LexA system. Confocal microscopy revealed that, in infected cells, TULV N protein and SUMO-1 colocalize at the perinuclear area providing further evidence for interaction between the two proteins. Neither endogenous nor transiently expressed SUMO-1 was found to be covalently linked to the N protein. Additional evidence that the interaction is non-covalent was obtained in immunoprecipitation experiments: N protein-specific antibodies precipitated SUMO-1 from TULV-infected Vero E6 cell lysate. By using a pepscan assay, two basic amino acid stretches in the N-terminal part of SUMO-1 were shown to be involved in the interaction.
Kaukinen
Pasi
P
Department of Virology, Haartman Institute, PO Box 21, University of Helsinki, FIN-00014, Helsinki, Finland. psi.kaukinen@helsinki.fi
Vaheri
Antti
A
Plyusnin
Alexander
A
D000595:Amino Acid Sequence; D000818:Animals; D001665:Binding Sites / Q000235:genetics; D002522:Chlorocebus aethiops; D018076:DNA, Complementary / Q000235:genetics; D006476:Orthohantavirus / Q000235:genetics / Q000378:metabolism*; D018613:Microscopy, Confocal; D008969:Molecular Sequence Data; D019590:Nucleocapsid Proteins / Q000235:genetics / Q000378:metabolism*; D010446:Peptide Fragments / Q000235:genetics / Q000378:metabolism; D011485:Protein Binding; D011994:Recombinant Proteins / Q000235:genetics / Q000378:metabolism; D025842:SUMO-1 Protein / Q000235:genetics / Q000378:metabolism*; D014162:Transfection; D020798:Two-Hybrid System Techniques; D014709:Vero Cells
D018076:DNA, Complementary; D019590:Nucleocapsid Proteins; D010446:Peptide Fragments; D011994:Recombinant Proteins; D025842:SUMO-1 Protein