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Cloning of ubiquitin-activating enzyme and ubiquitin-conjugating enzyme genes from Gracilaria lemaneiformis and their activity under heat shock.

Cloning of ubiquitin-activating enzyme and ubiquitin-conjugating enzyme genes from Gracilaria lemaneiformis and their activity under heat shock.

Authors :
Li, Guang-Qi
Zang, Xiao-Nan
Zhang, Xue-Cheng
Lu, Ning
Ding, Yan
Gong, Le
Chen, Wen-Chao
Source :
Gene. Mar2014, Vol. 538 Issue 1, p155-163. 9p.
Publication Year :
2014

Abstract

To study the response of Gracilaria lemaneiformis to heat stress, two key enzymes – ubiquitin-activating enzyme (E1) and ubiquitin-conjugating enzyme (E2) – of the Ubiquitin/26S proteasome pathway (UPP) were studied in three strains of G. lemaneiformis—wild type, heat-tolerant cultivar 981 and heat-tolerant cultivar 07-2. The full length DNA sequence of E1 contained only one exon. The open reading frame (ORF) sequence was 981 nucleotides encoding 326 amino acids, which contained conserved ATP binding sites (LYDRQIRLWGLE, ELAKNVLLAGV, LKEMN, VVCAI) and the ubiquitin-activating domains (VVCAI…LMTEAC, VFLDLGDEYSYQ, AIVGGMWGRE). The gene sequence of E2 contained four exons and three introns. The sum of the four exons gave an open reading frame sequence of 444 nucleotides encoding 147 amino acids, which contained a conserved ubiquitin-activating domain (GSICLDIL), ubiquitin-conjugating domains (RIYHPNIN, KVLLSICSLL, DDPLV) and ubiquitin-ligase (E3) recognition sites (KRI, YPF, WSP). Real-time-PCR analysis of transcription levels of E1 and E2 under heat shock conditions (28°C and 32°C) showed that in wild type, transcriptions of E1 and E2 were up-regulated at 28°C, while at 32°C, transcriptions of the two enzymes were below the normal level. In cultivar 981 and cultivar 07-2 of G. lemaneiformis, the transcription levels of the two enzymes were up-regulated at 32°C, and transcription level of cultivar 07-2 was even higher than that of cultivar 981. These results suggest that the UPP plays an important role in high temperature resistance of G. lemaneiformis and the bioactivity of UPP is directly related to the heat-resistant ability of G. lemaneiformis. [ABSTRACT FROM AUTHOR]

Details

Language :
English
ISSN :
03781119
Volume :
538
Issue :
1
Database :
Academic Search Index
Journal :
Gene
Publication Type :
Academic Journal
Accession number :
94405201
Full Text :
https://doi.org/10.1016/j.gene.2013.12.017