The vacuolar processing enzyme OsVPE1 is required for efficient glutelin processing in rice.
Rice (Oryza sativa L.) accumulates prolamines and glutelins as its major storage proteins. Glutelins are synthesized on rough endoplasmic reticulum as 57-kDa precursors; they are then sorted into protein storage vacuoles where they are processed into acidic and basic subunits. We report a novel rice glutelin mutant, W379, which accumulates higher levels of the 57-kDa glutelin precursor. Genetic analysis revealed that the W379 phenotype is controlled by a single recessive nuclear gene. Using a map-based cloning strategy, we identified this gene, OsVPE1, which is a homolog of the Arabidopsis betaVPE gene. OsVPE1 encodes a 497-amino-acid polypeptide. Nucleotide sequence analysis revealed a missense mutation in W379 that changes Cys269 to Gly. Like the wild-type protein, the mutant protein is sorted into vacuoles; however, the enzymatic activity of the mutant OsVPE1 is almost completely eliminated. Further, we show that OsVPE1 is incorrectly cleaved, resulting in a mature protein that is smaller than the wild-type mature protein. Taken together, these results demonstrate that OsVPE1 is a cysteine protease that plays a crucial role in the maturation of rice glutelins. Further, OsVPE1 Cys269 is a key residue for maintaining the Asn-specific cleavage activity of OsVPE1.
Wang
Yihua
Y
Jiangsu Plant Gene Engineering Research Center, Nanjing Agricultural University, China.
Zhu
Susong
S
Liu
Shijia
S
Jiang
Ling
L
Chen
Liangming
L
Ren
Yulong
Y
Han
Xiaohua
X
Liu
Feng
F
Ji
Sulan
S
Liu
Xi
X
Wan
Jianmin
J
D000595:Amino Acid Sequence; D001483:Base Sequence; D003001:Cloning, Molecular; D003546:Cysteine Endopeptidases / Q000235:genetics / Q000378:metabolism*; D005816:Genetic Complementation Test; D005983:Glutens / Q000378:metabolism*; D020125:Mutation, Missense; D012275:Oryza / Q000201:enzymology / Q000235:genetics*; D010940:Plant Proteins / Q000235:genetics / Q000378:metabolism*; D018749:RNA, Plant / Q000235:genetics; D017422:Sequence Analysis, DNA; D014617:Vacuoles / Q000201:enzymology*
D010940:Plant Proteins; D018749:RNA, Plant; D005983:Glutens; D003546:Cysteine Endopeptidases