Cloning and molecular characterization of a gene encoding late embryogenesis abundant protein from Pennisetum glaucum: protection against abiotic stresses.
Late embryogenesis abundant (LEA) protein family is a large protein family that protects other proteins from aggregation due to desiccation or osmotic stresses. A cDNA clone encoding a group 7 late embryogenesis abundant protein, termed PgLEA, was isolated from Pennisetum glaucum by screening a heat stress cDNA library. PgLEA cDNA encodes a 176 amino acid polypeptide with a predicted molecular mass of 19.21 kDa and an estimated isoelectric point of 7.77. PgLEA shares 70-74% sequence identity with other plant homologs. Phylogenetic analysis revealed that PgLEA is evolutionarily close to the LEA 7 group. Recombinant PgLEA protein expressed in Escherichia coli possessed in vitro chaperone activity and protected PgLEA-producing bacteria from damage caused by heat and salinity. Positive correlation existed between differentially up-regulated PgLEA transcript levels and the duration and intensity of different environmental stresses. In silico analysis of the promoter sequence of PgLEA revealed the presence of a distinct set of cis-elements and transcription factor binding sites. Transcript induction data, the presence of several putative stress-responsive transcription factor binding sites in the promoter region of PgLEA, the in vitro chaperone activity of this protein and its protective effect against heat and salt damage in E. coli suggest a role in conferring abiotic stress tolerance in plants.
International Centre for Genetic Engineering and Biotechnology (ICGEB), Aruna Asaf Ali Marg, New Delhi, India.
Palakolanu Sudhakar
PS
Reddy
Guda Maheedhar
GM
Reddy
Prachi
P
Pandey
Kottakota
K
Chandrasekhar
Malireddy K
MK
Reddy
D024506:5' Flanking Region; D000595:Amino Acid Sequence; D001483:Base Sequence; D003001:Cloning, Molecular; D004262:DNA Restriction Enzymes / Q000737:chemistry; D004795:Enzyme Stability; D004926:Escherichia coli / Q000235:genetics / Q000254:growth & development; D018506:Gene Expression Regulation, Plant; D006358:Hot Temperature; D008958:Models, Molecular; D008969:Molecular Sequence Data; D031782:Pennisetum / Q000235:genetics* / Q000254:growth & development / Q000502:physiology; D010802:Phylogeny; D010940:Plant Proteins / Q000096:biosynthesis / Q000737:chemistry / Q000235:genetics*; D011401:Promoter Regions, Genetic; D011489:Protein Denaturation; D017433:Protein Structure, Secondary; D017434:Protein Structure, Tertiary; D011994:Recombinant Proteins / Q000096:biosynthesis / Q000737:chemistry / Q000235:genetics; D055049:Salt Tolerance / Q000235:genetics; D017422:Sequence Analysis, DNA; D013312:Stress, Physiological; D014158:Transcription, Genetic; D015854:Up-Regulation
D010940:Plant Proteins; D011994:Recombinant Proteins; C066246:late embryogenesis abundant protein, plant; D004262:DNA Restriction Enzymes