Association of the nucleocapsid protein of the Seoul and Hantaan hantaviruses with small ubiquitin-like modifier-1-related molecules.
We performed yeast two-hybrid screening of a human kidney cell cDNA library to study the biological role of the hantavirus nucleocapsid protein (NP). We found that Seoul virus (SEOV) and Hantaan virus (HTNV) NPs were associated with small ubiquitin-like modifier (SUMO)-1-interacting proteins PIAS1, PIASxbeta, HIPK2, CHD3, and TTRAP, which interacted with the SUMO-1 conjugating enzyme (Ubc-9) and SUMO-1 in the yeast two-hybrid assay. Interactions between the HIPK2, CHD3, and TTRAP proteins and SEOV NP were also shown in a mammalian two-hybrid assay. However, there was no interaction between PIAS proteins and NP, which was probably due to the inhibitory effect of PIAS on transcription in the mammalian two-hybrid assay. Nevertheless, a co-expression experiment suggested the existence of a PIAS-NP interaction in the cytoplasm. The region spanning amino acids 100-125 of SEOV NP, which represents a critical region for NP-NP polymerization, was found to be responsible for the interaction with SUMO-1-related molecules in both the yeast and mammalian two-hybrid assays. These results add to the information on interactions of hantavirus NP and host cellular proteins.
Lee
Byoung-Hee
BH
Institute for Animal Experimentation, Graduate School of Medicine, Hokkaido University, Kita-15, Nishi-7, Kita-ku, Sapporo 060-8638, Japan.
Yoshimatsu
Kumiko
K
Maeda
Akihiko
A
Ochiai
Kazuhiko
K
Morimatsu
Masami
M
Araki
Koichi
K
Ogino
Michiko
M
Morikawa
Shigeru
S
Arikawa
Jiro
J
D000818:Animals; D036022:Capsid Proteins / Q000378:metabolism*; D002460:Cell Line; D002522:Chlorocebus aethiops; D003593:Cytoplasm / Q000378:metabolism; D015723:Gene Library; D018063:Hantaan virus / Q000378:metabolism*; D006367:HeLa Cells; D006801:Humans; D019590:Nucleocapsid Proteins / Q000378:metabolism*; D050828:Protein Inhibitors of Activated STAT; D011506:Proteins / Q000378:metabolism*; D025842:SUMO-1 Protein / Q000378:metabolism*; D029263:Seoul virus / Q000378:metabolism*; D044763:Ubiquitin-Conjugating Enzymes / Q000378:metabolism; D014709:Vero Cells; D014758:Viral Core Proteins / Q000378:metabolism*; D000098843:Ubiquitin-Conjugating Enzyme UBC9
D036022:Capsid Proteins; D019590:Nucleocapsid Proteins; D050828:Protein Inhibitors of Activated STAT; D011506:Proteins; D025842:SUMO-1 Protein; D014758:Viral Core Proteins; C111782:nucleocapsid protein, Hantaan virus; D044763:Ubiquitin-Conjugating Enzymes; D000098843:Ubiquitin-Conjugating Enzyme UBC9