Molecular characterization of rice hsp101: complementation of yeast hsp104 mutation by disaggregation of protein granules and differential expression in indica and japonica rice types.
HSP100 protein is an important component of the heat-shock response in diverse organisms. Using specific primers based on cDNA sequence, rice hsp101 gene was PCR-amplified and sequenced. Southern analysis revealed that there appears to be a single gene per haploid genome coding for HSP101 protein in rice. Northern analysis showed that expression of hsp101 transcript is strictly heat-inducible and induction is transient in nature. In the temperature regime tested, 45 degrees C treatment to intact rice seedlings for 2 h showed maximal levels of hsp101 mRNA. Rice full-length hsp101 cDNA complemented yeast mutant disrupted for its own hsp104 gene by insertional mutagenesis, with efficacy that was comparable with Arabidopsis hsp101 cDNA. Electron micrographic evidence suggested that rice hsp101 cDNA in yeast is active in re-solubilizing the stress-induced protein granules in the post-stress recovery period. Rice hsp101 cDNA expression in hsp104 deficient yeast also caused recovery in tolerance against arsenite. Western analyses showed that this protein is expressed more rapidly during the stress period and retained for longer duration in the post-stress recovery period in japonica rice as compared to indica rice types. This is the first report wherein plant HSP100 protein expression is correlated to disappearance of protein granules in the yeast cells and distinct rice type-dependent protein expression patterns are reported.
Agarwal
Manu
M
Department of Plant Molecular Biology, University of Delhi South Campus, Benito Juarez Road, Dhaula Kuan, New Delhi-110021, India.
Sahi
Chandan
C
Katiyar-Agarwal
Surekha
S
Agarwal
Sangeeta
S
Young
Todd
T
Gallie
Daniel R
DR
Sharma
Vishva Mitra
VM
Ganesan
K
K
Grover
Anil
A
D018053:Arsenites / Q000494:pharmacology; D015139:Blotting, Southern; D015153:Blotting, Western; D002455:Cell Division / Q000187:drug effects / Q000235:genetics; D003001:Cloning, Molecular; D018744:DNA, Plant / Q000737:chemistry / Q000235:genetics; D018506:Gene Expression Regulation, Plant; D017343:Genes, Plant / Q000235:genetics; D005816:Genetic Complementation Test; D006360:Heat-Shock Proteins / Q000235:genetics; D008969:Molecular Sequence Data; D009154:Mutation; D012275:Oryza / Q000235:genetics*; D010940:Plant Proteins / Q000235:genetics* / Q000378:metabolism; D012333:RNA, Messenger / Q000235:genetics / Q000378:metabolism; D012441:Saccharomyces cerevisiae / Q000235:genetics / Q000254:growth & development / Q000648:ultrastructure; D029701:Saccharomyces cerevisiae Proteins / Q000235:genetics; D017422:Sequence Analysis, DNA; D017670:Sodium Compounds / Q000494:pharmacology; D013045:Species Specificity; D013696:Temperature; D014157:Transcription Factors / Q000235:genetics* / Q000378:metabolism
D018053:Arsenites; D018744:DNA, Plant; C115569:HSP101 protein, plant; D006360:Heat-Shock Proteins; D010940:Plant Proteins; D012333:RNA, Messenger; D029701:Saccharomyces cerevisiae Proteins; D017670:Sodium Compounds; D014157:Transcription Factors; C070972:HsP104 protein, S cerevisiae; C017947:sodium arsenite