Primary structure and differential expression of beta-amylase in normal and mutant barleys.
The primary structure of barley endosperm beta-amylase, an enzyme which catalyses the liberation of maltose from 1,4-alpha-D-glucans, has been deduced from the nucleotide sequence of a cloned full-length cDNA. The mRNA is 1754 nucleotides long [excluding the poly(A) tail] and codes for a polypeptide of 535 amino acids with a relative molecular mass of 59,663. The deduced amino acid sequence was compared with the sequences of ten peptides obtained from the purified enzyme and unambiguous identification was obtained. The N-terminal region of the deduced sequence was identical to a 12-residue cyanogen-bromide-peptide sequence, indicating that beta-amylase is synthesized as the mature protein. A graphic matrix homology plot shows four glycine-rich repeats, each of 11 residues, preceding the C-terminus. Southern blotting of genomic DNA demonstrates that beta-amylase is encoded by a small gene family, while cDNA sequence analysis indicates the presence of at least two types of mRNA in the endosperm. Dot and northern blot analysis show that Hiproly barley contains greatly increased levels of beta-amylase mRNA compared to the normal cultivar Sundance, whereas Risø mutant 1508 contains only trace amounts. These results correlate well with the deposition of beta-amylase during endosperm development in these lines. Low but similar amounts of beta-amylase mRNAs sequences were detected in leaves and shoots from normal and mutant barleys, demonstrating that the mutant lys3a (1508) and lysl (Hiproly) genes do not affect the expression of beta-amylase in these tissues.
D000595:Amino Acid Sequence; D000681:Amylases / Q000032:analysis*; D001483:Base Sequence; D004247:DNA / Q000032:analysis; D005786:Gene Expression Regulation; D005796:Genes; D001467:Hordeum; D008969:Molecular Sequence Data; D009154:Mutation; D010449:Peptide Mapping; D010944:Plants / Q000201:enzymology* / Q000235:genetics; D001614:beta-Amylase / Q000032:analysis* / Q000235:genetics
D004247:DNA; D000681:Amylases; D001614:beta-Amylase