Construction and functional characterization of double and triple mutants of parallel <img src='http://www.niscair.res.in/jinfo/Beta.gif' border=0>-bulge of ubiquitin
IR@NISCAIR: CSIR-NISCAIR, New Delhi - ONLINE PERIODICALS REPOSITORY (NOPR)
View Archive Info| Field | Value | |
| Creator |
Sharma, Mrinal
Prabha, C Ratna |
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| Date |
2011-11-30T09:37:15Z
2011-11-30T09:37:15Z 2011-12 |
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| Identifier |
0975-1009 (Online); 0019-5189 (Print)
http://hdl.handle.net/123456789/13098 |
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| Description |
919-924
Ubiquitin, a small eukaryotic protein serving as a post-translational modification on many important proteins, plays central role in cellular homeostasis and cell cycle regulation. Ubiquitin features two <img src='http://www.niscair.res.in/jinfo/Beta.gif' border=0>-bulges, the second <img src='http://www.niscair.res.in/jinfo/Beta.gif' border=0>-bulge, located at the C-terminal region of the protein along with type II turn, holds 3 residues Glu64(1), Ser65(2) and Gln2(X). Percent frequency of occurrence of such a sequence in parallel <img src='http://www.niscair.res.in/jinfo/Beta.gif' border=0>-bulge is very low. However, the sequence and structure have been conserved in ubiquitin through out the evolution. Present study involves replacement of residues in unusual <img src='http://www.niscair.res.in/jinfo/Beta.gif' border=0>-bulge of ubiquitin by introducing mutations in combination through site directed mutagenesis, generating double and triple mutants and their functional characterization. Mutant ubiquitins cloned in yeast expression vector YEp96 tested for growth profile, viability assay and heat stress complementation study have revealed significant decrease in growth rate, loss of viability and non-complementation of heat sensitive phenotype with UbE64G-S65D and UbQ2N-E64G-S65D mutations. However, UbQ2N-S65D did not show any negative effects in the above assays. Present results show that, replacement of residues in <img src='http://www.niscair.res.in/jinfo/Beta.gif' border=0>-bulge of ubiquitin exerts severe effects on growth and viability in <i style="">Saccharomyces cerevisiae </i>due to functional failure of the mutant ubiquitins UbE64G-S65D and UbQ2N-E64G-S65D. |
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| Language |
en_US
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| Publisher |
NISCAIR-CSIR, India
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| Rights |
<img src='http://nopr.niscair.res.in/image/cc-license-sml.png'> <a href='http://creativecommons.org/licenses/by-nc-nd/2.5/in' target='_blank'>CC Attribution-Noncommercial-No Derivative Works 2.5 India</a>
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| Source |
IJEB Vol.49(12) [December 2011]
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| Subject |
<img src='http://www.niscair.res.in/jinfo/Beta.gif' border=0>-Bulge
Cancer Ubiquitin Ubiquitin proteasome system (UPS) Virus |
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| Title |
Construction and functional characterization of double and triple mutants of parallel <img src='http://www.niscair.res.in/jinfo/Beta.gif' border=0>-bulge of ubiquitin
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| Type |
Article
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