Functional study of hot pepper 26S proteasome subunit RPN7 induced by Tobacco mosaic virus from nuclear proteome analysis.
Two-dimensional gel electrophoresis (2-DE) was applied for the screening of Tobacco mosaic virus (TMV)-induced hot pepper (Capsicum annuum cv. Bugang) nuclear proteins. From differentially expressed protein spots, we acquired the matched peptide mass fingerprint (PMF) data, analyzed by MALDI-TOF MS, from the non-redundant hot pepper EST protein FASTA database using the VEMS 2.0 software. Among six identified nuclear proteins, the hot pepper 26S proteasome subunit RPN7 (CaRPN7) was subjected to further study. The level of CaRPN7 mRNA was specifically increased during incompatible TMV-P(0) interaction, but not during compatible TMV-P(1.2) interaction. When CaRPN7::GFP fusion protein was targeted in onion cells, the nuclei had been broken into pieces. In the hot pepper leaves, cell death was exacerbated and genomic DNA laddering was induced by Agrobacterium-mediated transient overexpression of CaPRN7. Thus, this report presents that the TMV-induced CaRPN7 may be involved in programmed cell death (PCD) in the hot pepper plant.
Lee
Boo-Ja
BJ
School of Life Sciences and Biotechnology, Korea University, Seoul 136-701, Republic of Korea.
Kwon
Sun Jae
SJ
Kim
Sung-Kyu
SK
Kim
Ki-Jeong
KJ
Park
Chang-Jin
CJ
Kim
Young-Jin
YJ
Park
Ohkmae K
OK
Paek
Kyung-Hee
KH
D000595:Amino Acid Sequence; D017209:Apoptosis; D002212:Capsicum / Q000378:metabolism* / Q000821:virology; D002467:Cell Nucleus / Q000378:metabolism*; D015180:Electrophoresis, Gel, Two-Dimensional; D018506:Gene Expression Regulation, Plant; D008969:Molecular Sequence Data; D019697:Onions / Q000378:metabolism / Q000821:virology; D018515:Plant Leaves / Q000378:metabolism / Q000821:virology; D010940:Plant Proteins / Q000378:metabolism*; D046988:Proteasome Endopeptidase Complex / Q000096:biosynthesis / Q000378:metabolism*; D021122:Protein Subunits / Q000378:metabolism; D020543:Proteome / Q000378:metabolism*; D012231:Rhizobium / Q000235:genetics; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization; D014027:Tobacco Mosaic Virus / Q000502:physiology*
D010940:Plant Proteins; D021122:Protein Subunits; D020543:Proteome; D046988:Proteasome Endopeptidase Complex; C061553:ATP dependent 26S protease