Analysis of regulatory elements of the promoter and the 3' untranslated region of the maize Hrgp gene coding for a cell wall protein.
Hydroxyproline-rich glycoproteins (HRGP) are structural components of the plant cell wall. Hrgp genes from maize and related species have a conserved 500 bp sequence in the 5'-flanking region, and all Hrgp genes from monocots have an intron located in the 3' untranslated region. To study the role of these conserved regions, several deletions of the Hrgp gene were fused to the beta-glucuronidase ( GUS) gene and used to transform maize tissues by particle bombardment. The overall pattern of GUS activity directed by sequential deletions of the Hrgp promoter was different in embryos and young shoots. In embryos, the activity of the full-length Hrgp promoter was in the same range as that of the p35SI promoter construct, based on the strong 35S promoter, whereas in the fast-growing young shoots it was 20 times higher. A putative silencer element specific for young shoots was found in the -1,076/-700 promoter region. Other major cis elements for Hrgp expression are probably located in the regions spanning -699/-510 and -297/-160. Sequences close to the initial ATG and mRNA leader were also important since deletion of the region -52/+16 caused a 75% reduction in promoter activity. The presence of the Hrgp intron in the 3' untranslated region changed the levels of GUS activity directed by the Hrgp and the 35S promoters. This pattern of activity was complex, and was dependent on the promoter and cell type analysed.
Menossi
M
M
Centro de Biologia Molecular e Engenharia Genética, Universidade Estadual de Campinas (UNICAMP), CxP 6010, CEP 13083-970, Campinas SP, Brazil. menossi@unicamp.br
Rabaneda
F
F
Puigdomènech
P
P
Martínez-Izquierdo
J A
JA
D020413:3' Untranslated Regions; D001483:Base Sequence; D019470:Biolistics; D018744:DNA, Plant; D006023:Glycoproteins / Q000235:genetics*; D008969:Molecular Sequence Data; D010940:Plant Proteins / Q000235:genetics*; D011401:Promoter Regions, Genetic; D012045:Regulatory Sequences, Nucleic Acid; D017384:Sequence Deletion; D003313:Zea mays / Q000235:genetics*
D020413:3' Untranslated Regions; D018744:DNA, Plant; D006023:Glycoproteins; D010940:Plant Proteins; C005488:extensin protein, plant