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| Title: Genome-wide analysis of rice ClpB/HSP100 , ClpC and ClpD genes .
| | Author: Singh A Singh U Mittal D Grover A | | Journal: BMC Genomics Citation: V : 11 P : 95 Year: 2010 Type: MEDLINE | | Literature: oryza Field: abstract Doc ID: pub20141629 Accession (PMID): 20141629 | | Abstract: BACKGROUND : ClpB-cyt/HSP100 protein acts as chaperone , mediating disaggregation of denatured proteins .
Previous studies have shown that ClpB-cyt/HSP100 gene belongs to the group class I Clp ATPase proteins and ClpB-cyt/HSP100 transcript is regulated by heat stress and developmental cues .
RESULTS : Nine ORFs were noted to constitute rice class I Clp ATPases in the following manner : 3 ClpB proteins ( ClpB-cyt , Os05g44340 ; ClpB-m , Os02g08490 ; ClpB-c , Os03g31300 ) , 4 ClpC proteins ( ClpC1 , Os04g32560 ; ClpC2 , Os12g12580 ; ClpC3 , Os11g16590 ; ClpC4 , Os11g16770 ) and 2 ClpD proteins ( ClpD1 , Os02g32520 ; ClpD2 , Os04g33210 ) .
Using the respective signal sequences cloned upstream to GFP/CFP reporter proteins and transient expression studies with onion epidermal cells , evidence is provided that rice ClpB-m and Clp-c proteins are indeed localized to their respective cell locations mitochondria and chloroplasts , respectively .
Associated with their diverse cell locations , domain structures of OsClpB-c , OsClpB-m and OsClpB-cyt proteins are noted to possess a high-level conservation .
OsClpB-cyt transcript is shown to be enriched at milk and dough stages of seed development .
While expression of OsClpB-m was significantly less as compared to its cytoplasmic and chloroplastic counterparts in different it issues , this transcript showed highest heat-induced expression amongst the 3 ClpB proteins .
OsClpC1 and OsClpC2 are predicted to be chloroplast-localized as is the case with all known plant ClpC proteins .
However , the fact that OsClpC3 protein appears mitochondrial/chloroplastic with equal probability and OsClpC4 a plasma membrane protein reflects functional diversity of this class .
Different class I Clp ATPase transcripts were noted to be cross-induced by a host of different abiotic stress conditions .
Complementation assays of Deltahsp104 mutant yeast cells showed that OsClpB-cyt , OsClpB-m , OsClpC1 and OsClpD1 have significantly positive effects .
Remarkably , OsClpD1 gene imparted appreciably high level tolerance to the mutant yeast cells .
CONCLUSIONS : Rice class I Clp ATPase gene family is constituted of 9 members .
Of these 9 , only 3 belonging to ClpB group are heat stress regulated .
Distribution of ClpB proteins to different cell organelles indicates that their functioning might be critical in different cell locations .
From the complementation assays , OsClpD1 appears to be more effective than OsClpB-cyt protein in rescuing the thermosensitive defect of the yeast ScDeltahsp104 mutant cells .
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[ Sen. 3, subscore: 1.00 ]: RESULTS : Nine ORFs were noted to constitute rice class I Clp ATPases in the following manner : 3 ClpB proteins ( ClpB-cyt , Os05g44340 ; ClpB-m , Os02g08490 ; ClpB-c , Os03g31300 ) , 4 ClpC proteins ( ClpC1 , Os04g32560 ; ClpC2 , Os12g12580 ; ClpC3 , Os11g16590 ; ClpC4 , Os11g16770 ) and 2 ClpD proteins ( ClpD1 , Os02g32520 ; ClpD2 , Os04g33210 ) .
[ Sen. 4, subscore: 1.00 ]: Using the respective signal sequences cloned upstream to GFP/CFP reporter proteins and transient expression studies with onion epidermal cells , evidence is provided that rice ClpB-m and Clp-c proteins are indeed localized to their respective cell locations mitochondria and chloroplasts , respectively .
| | Supplemental links/files: reference in endnote online text related articles pubmed citation | |
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| Title: OsHsfA2c and OsHsfB4b are involved in the transcriptional regulation of cytoplasmic OsClpB ( Hsp100 ) gene in rice ( Oryza sativa L ) .
| | Author: Singh A Mittal D Lavania D Agarwal M Mishra RC Grover A | | Journal: Cell Stress Chaperones Citation: V : 17 P : 243-54 Year: 2012 Type: In-Process | | Literature: oryza Field: abstract Doc ID: pub22147560 Accession (PMID): 22147560 | | Abstract: ClpB-cytoplasmic ( ClpB-cyt ) /Hsp100 is an important chaperone protein in rice .
Cellular expression of OsClpB-cyt transcript is governed by heat stress , metal stress , and developmental cues .
Transgenic rice plants produced with 2 kb OsClpB-cyt promoter driving Gus reporter gene showed heat and metal-regulated Gus expression in vegetative it issues and constitutive Gus expression in calli , flowering it issues , and embryonal half of seeds .
Rice seedlings regenerated with OsClpB-cyt promoter fragment with deletion of its canonical heat shock element sequence ( HSE ( -273 to -280 ) ) showed not only heat shock inducibility of Gus transcript/protein but also constitutive expression of Gus in vegetative it issues .
It thus emerges that the only classical HSE present in OsClpB-cyt promoter is involved in repressing expression of OsClpB-cyt transcript under unstressed control conditions .
Yeast one-hybrid assays suggested that OsHsfA2c specifically interacts with OsClpB-cyt promoter .
OsHsfA2c also showed binding with OsClpB-cyt and OsHsfB4b showed binding with OsClpB-cyt ; notably , interaction of OsHsfB4b was seen for all three OsClpB/Hsp100 protein isoforms ( ie , ClpB-cytoplasmic , ClpB-mitochondrial , and ClpB-chloroplastic ) .
Furthermore , OsHsfB4b showed interaction with OsHsfA2c .
This study suggests that OsHsfA2c may play a role as transcriptional activator and that OsHsfB4b is an important part of this heat shock responsive circuitry .
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[ Sen. 7, subscore: 1.00 ]: OsHsfA2c also showed binding with OsClpB-cyt and OsHsfB4b showed binding with OsClpB-cyt ; notably , interaction of OsHsfB4b was seen for all three OsClpB/Hsp100 protein isoforms ( ie , ClpB-cytoplasmic , ClpB-mitochondrial , and ClpB-chloroplastic ) .
| | Supplemental links/files: reference in endnote online text related articles pubmed citation | |
