Identification and functional analysis of salmon annexin 1 induced by a virus infection in a fish cell line.
In this study, we investigated changes in protein expression of fish cells induced by infection of infectious pancreatic necrosis virus (IPNV) using two-dimensional electrophoresis and matrix-assisted laser desorption-time of flight proton motive force analysis and identified a novel type of salmon annexin 1 that is induced in fish cells by infection with IPNV. Northern blotting showed that this annexin is overexpressed in IPNV-infected cells compared to control cells, and further analysis revealed that it has a 1,509-bp full-length cDNA sequence with an open reading frame encoding 339 amino acids (GenBank accession no. AY944135). Amino acid sequence analysis revealed that this protein belongs to the annexin 1 subfamily. By applying RNA interference, the mRNA levels of salmon annexin 1 were suppressed and, under these conditions, apoptosis of IPNV-infected cells was significantly increased. While small interfering RNA (siRNA) treatment did not affect the levels of the viral proteins significantly until 10 h postinfection, it reduced the titer of extracellular virus to 25% of that of a scrambled siRNA-treated control. These data provide evidence of an antiapoptotic function for salmon annexin 1 that is important for IPNV growth in cultured cells.
Hwang
Hyun Jin
HJ
Graduate School of Biotechnology and Institute of Life Science and Resources, Kyung Hee University, Yongin 449-701, Korea.
Moon
Chang Hoon
CH
Kim
Han Geun
HG
Kim
Joo Yun
JY
Lee
Jung Min
JM
Park
Jeong Woo
JW
Chung
Dae Kyun
DK
D000595:Amino Acid Sequence; D000818:Animals; D017302:Annexins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism / Q000494:pharmacology; D017209:Apoptosis / Q000187:drug effects; D002478:Cells, Cultured; D015180:Electrophoresis, Gel, Two-Dimensional; D005786:Gene Expression Regulation; D018061:Infectious pancreatic necrosis virus / Q000472:pathogenicity*; D008969:Molecular Sequence Data; D010802:Phylogeny; D012333:RNA, Messenger / Q000235:genetics / Q000378:metabolism; D034741:RNA, Small Interfering / Q000235:genetics / Q000378:metabolism; D012474:Salmon / Q000821:virology*; D017422:Sequence Analysis, DNA; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization
D017302:Annexins; D012333:RNA, Messenger; D034741:RNA, Small Interfering