title: Identification, characterization, and analysis of cDNA and genomic sequences encoding two different small heat shock proteins in Hordeum vulgare.
abstract: In vitro translation of mRNAs prepared from barley (Hordeum vulgare) seedlings (cv. Onice) exposed at 40 degrees C directed the synthesis of major heat shock proteins (HSPs) with molecular masses of 80-90, 70, 42 and 16-22 kDa. A cDNA library prepared from the 40 degrees C mRNAs and screened by differential hybridization led to the isolation of heat shock specific sequences. One of these (Hv hsp18) was confirmed by hybrid-arrested and hybrid-released translation as encoding for an 18-kDa HSP. The barley hsp18 sequence has an open reading frame encoding a 160 amino acid residue 18-kDa protein that is 63% identical to wheat 16.9-kDa HSP (clone C5-8), 54% identical to soybean (Glycine max) 17.5-kDa HSP, and 49% identical to Arabidopsis thaliana 17.6-kDa HSP. Lower similarities were found with class II plant small HSPs such as soybean 17.9-kDa HSP (27%), Pisum sativum 17.7-kDa HSP (30%), wheat (Triticum aestivum) 17.3-kDa HSP (clone Ta hsp 17.3) (30%), and with animal small HSPs and alpha-crystallins. The Hv hsp18 sequence was used to pick up Hv hsp17 genomic sequence encoding for another class I 17-kDa HSP. By computer analysis of the nucleotide sequence the TATA box, two heat shock promoter elements, a metal-ion response element, and the polyadenylation signals were identified. Barley HSP18 has an additional cysteine-rich region when compared with HSP17 mapping at the carboxy terminal end.
authors.affiliation: Department of Evolutionary Biology, University of Parma, Italy.
authors.forename: N
authors.initials: N
authors.lastname: Marmiroli
authors.forename: A
authors.initials: A
authors.lastname: Pavesi
authors.forename: G
authors.initials: G
authors.lastname: Di Cola
authors.forename: H
authors.initials: H
authors.lastname: Hartings
authors.forename: G
authors.initials: G
authors.lastname: Raho
authors.forename: M R
authors.initials: MR
authors.lastname: Conte
authors.forename: C
authors.initials: C
authors.lastname: Perrotta
mesh_terms: D000595:Amino Acid Sequence; D001483:Base Sequence; D003001:Cloning, Molecular; D018076:DNA, Complementary / Q000235:genetics*; D017343:Genes, Plant; D006360:Heat-Shock Proteins / Q000737:chemistry / Q000235:genetics*; D001467:Hordeum / Q000235:genetics*; D008969:Molecular Sequence Data; D008970:Molecular Weight; D010940:Plant Proteins / Q000737:chemistry / Q000235:genetics*; D012333:RNA, Messenger / Q000235:genetics; D015183:Restriction Mapping; D017386:Sequence Homology, Amino Acid
chemical_list: D018076:DNA, Complementary; C489905:HSP18 protein, plant; D006360:Heat-Shock Proteins; D010940:Plant Proteins; D012333:RNA, Messenger