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Journal of Zhejiang University SCIENCE B 2014 Vol.15 No.7 P.624-637


Molecular identification and interaction assay of the gene (OsUbc13) encoding a ubiquitin-conjugating enzyme in rice*

Author(s):  Ya Wang1,2, Meng-yun Xu1, Jian-ping Liu1, Mu-gui Wang1, Hai-qing Yin2, Ju-min Tu1

Affiliation(s):  1. Institute of Crop Science, College of Agriculture and Biotechnology, Zhejiang University, Hangzhou 310058, China; more

Corresponding email(s):   jtu@zju.edu.cn

Key Words:  Ubc13, DNA damage tolerance, Oryza sativa , Real-time quantitative PCR, Yeast two-hybrid

Ya Wang, Meng-yun Xu, Jian-ping Liu, Mu-gui Wang, Hai-qing Yin, Ju-min Tu. Molecular identification and interaction assay of the gene (OsUbc13) encoding a ubiquitin-conjugating enzyme in rice[J]. Journal of Zhejiang University Science B, 2014, 15(7): 624-637.

@article{title="Molecular identification and interaction assay of the gene (OsUbc13) encoding a ubiquitin-conjugating enzyme in rice",
author="Ya Wang, Meng-yun Xu, Jian-ping Liu, Mu-gui Wang, Hai-qing Yin, Ju-min Tu",
journal="Journal of Zhejiang University Science B",
publisher="Zhejiang University Press & Springer",

%0 Journal Article
%T Molecular identification and interaction assay of the gene (OsUbc13) encoding a ubiquitin-conjugating enzyme in rice
%A Ya Wang
%A Meng-yun Xu
%A Jian-ping Liu
%A Mu-gui Wang
%A Hai-qing Yin
%A Ju-min Tu
%J Journal of Zhejiang University SCIENCE B
%V 15
%N 7
%P 624-637
%@ 1673-1581
%D 2014
%I Zhejiang University Press & Springer
%DOI 10.1631/jzus.B1300273

T1 - Molecular identification and interaction assay of the gene (OsUbc13) encoding a ubiquitin-conjugating enzyme in rice
A1 - Ya Wang
A1 - Meng-yun Xu
A1 - Jian-ping Liu
A1 - Mu-gui Wang
A1 - Hai-qing Yin
A1 - Ju-min Tu
J0 - Journal of Zhejiang University Science B
VL - 15
IS - 7
SP - 624
EP - 637
%@ 1673-1581
Y1 - 2014
PB - Zhejiang University Press & Springer
ER -
DOI - 10.1631/jzus.B1300273

The ubiquitin (Ub)-conjugating enzyme, ubc13, has been known to be involved in error-free DNA damage tolerance (or post-replication repair) via catalyzing Lys63-linked polyubiquitin chains formation together with a Ubc variant. However, its functions remain largely unknown in plant species, especially in monocotyledons. In this study, we cloned a Ub-conjugating enzyme, Osubc13, that shares the conserved domain of Ubc with Atubc13B in Oryza sativa L., which encodes a protein of 153 amino acids; the deduced sequence shares high similarities with other homologs. Real-time quantitative polymerase chain reaction (PCR) indicated that Osubc13 transcripts could be detected in all tissues examined, and the expression level was higher in palea, pistil, stamen, and leaf, and lower in root, stem, and lemma; the expression of Osubc13 was induced by low temperature, methylmethane sulfate (MMS), and H2O2, but repressed by mannitol, abscisic acid (ABA), and NaCl. Osubc13 was probably localized in the plasma and nuclear membranes. About 20 proteins, which are responsible for the positive yeast two-hybrid interaction of Osubc13, were identified. These include the confirmed OsVDAC (correlated with apoptosis), OsMADS1 (important for development of floral organs), OsB22EL8 (related to reactive oxygen species (ROS) scavenging and DNA protection), and OsCROC-1 (required for formation of Lys63 polyubiquitylation and error-free DNA damage tolerance). The molecular characterization provides a foundation for the functional study of Osubc13.


重要结论:OsUbc13编码具有153个氨基酸的蛋白质,其推断的氨基酸序列与其它同源序列具有很高的相似性;该基因在水稻各组织中均有表达,其中内稃、雌蕊、雄蕊和叶片中的表达量较高,而根、茎和外稃中的表达量较低; 低温、甲基磺酸甲酯(MMS)和过氧化氢(H2O2)胁迫处理使胚性愈伤中OsUbc13的表达量显著上调,甘露醇、脱落酸(ABA)和氯化钠(NaCl)胁迫则使愈伤组织中该基因的表达量降低;OsUbc13与绿色荧光蛋白(GFP)的融合蛋白表达于质膜和核膜处;酵母双杂交结果表明约有20个蛋白可能与OsUbc13存在相互作用,其中OsVDAC(与细胞凋亡有关)、OsMADS1(与花器官发育有关)、OsB22EL8(与活性氧清除及DNA保护有关)和OsCROC-1(为Lys63聚合泛素链形成及运行无误性DNA损伤耐受机制所必需)四个蛋白经验证确与OsUbc13互作。


Darkslateblue:Affiliate; Royal Blue:Author; Turquoise:Article


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