Abstract
Deletion analysis of the promoter sequence of the tobacco gene encoding a pollen-specific polygalacturonase (Npg1) revealed several motifs related to the LAT52/56 and LAT56/59 boxes (Twell et al. 1991) and a 14 bp element with homology to the maize polygalacturonase promoter (Allen and Lonsdale 1993), termed the PG box. Deletion analysis also revealed a complex pattern of domains which increase or reduce promoter activity. In particular, deleton of the LAT52/56 and LAT56/59 box motifs resulted in significant reductions in expression giving weight to the idea that such sequences function as transcriptional positive regulatory elements. Deletion of sequences immediately 3′ to the LAT52/56 and LAT56/59 motifs fully restore promoter function, suggesting the presence of suppressor elements associated with these positive regulatory elements. Analysis of the deletion series allowed the promoter to be divided into four domains: a modulation domain (-744 to -362), a basic promoter (-362), a core promoter (-267) and a minimal promoter (-182). Two sequence elements, Eh-1 (22 bp) and Eh-2 (28 bp), were identified between -362 and -267. Deletion of these two unrelated sequence elements reduces the activity of the basic promoter to that of the core promoter, a level of activity which is retained by the minimal promoter. A background level of activity is reached when the promoter is reduced to -86. Therefore, the sequence between -182 and -86, which contains two LAT52/56 motifs and the PG box, contains sufficient information to direct efficient gene expression.
Article PDF
Similar content being viewed by others
Avoid common mistakes on your manuscript.
References
Albani D, Sardana R, Robert LS, Altosaar I, Arnison PG, Fabijanski SF (1992) A Brassica napus gene family which shows sequence similarity to ascorbate oxidase is expressed in developing pollen. Molecular characterization and analysis of promoter activity in transgenic tobacco plants. Plant J 2:331–342
Allen RL, Lonsdale DM (1993) Molecular characterisation of one of the maize polygalacturonase gene family members, which are expressed during late pollen development. Plant J 3: 261–271
Fabijanski SF, Albani D, Robert LS, Arnison PG (1992) Characterization of genes expressed during the development of Brassica napus pollen. In Vitro Cell Dev Biol 280:46–52
Green PG, Yong M-H, Cuozzo M, Kano-Murakami Y, Silverstein P, Chua N-H (1988) Binding site requirements for pea nuclear protein factor GT-1 correlate with sequences required for light-dependent transcriptional activation of the rbcS-3A gene. EMBO J 7:4035–4044
Hamilton DA, Bashe DM, Stinson JR, Mascarenhas JP (1989) Characterization of a pollen-specific genomic clone from maize. Sex Plant Reprod 2:208–212
Hanson DD, Hamilton DA, Travis JL, Bashe DM, Mascarenhas JP (1989) Characterization of a pollen-specific cDNA clone from Zea mays and its expression. Plant Cell 1:173–179
Henikoff S (1987) Unidirectional digestion with exonuclease III in DNA sequence analysis. Methods Enzymol 155:156–165
Jefferson RA (1987) Assaying chimeric genes in plants: the GUS gene fusion system. Plant Mol Biol Rep 5:387–405
Joshi CP (1987) An inspection of the domain between putative TATA box and translation start site in 79 plant genes. Nucleic Acids Res 15:6643–6653
Lonsdale DM, Hodge TP, Stoehr PJ (1988) Analysis of the genome structure of plant mitochondria. In: Weissbach A, Weissbach H (eds) Methods for plant molecular biology. Academic Press, New York, pp 275–292
Lonsdale DM, Onde S, Cuming A (1990) Transient expression of exogenous DNA in intact, viable wheat embryos following particle bombardment. J Exp Bot 41:1161–1165
Murashige T, Skoog F (1962) A revised medium for rapid growth and bio assays with tobacco tissue cultures. Physiol Plant 15:473–497
Ow DW, Wood KV, DeLuca M, deWet JR, Helinski DR, Howell SH (1986) Transient and stable expression of the firefly luciferase gene in plant cells and transgenic plants. Science 234:856–859
Rogers HJ, Harvey A, Lonsdale DM (1992) Isolation and characterization of a tobacco gene with homology to pectate lyase which is specifically expressed during microsporogenesis. Plant Mol Biol 20:493–502
Snedecor GW (1967) Statistical methods applied to experiments in agriculture and biology. Iowa State University Press, Ames, Iowa
Tebbutt SJ, Rogers HJ, Lonsdale DM (1994) Characterization of a tobacco gene encoding a pollen-specific polygalacturonase. Plant Mol Biol 25:283–297
Thangavelu M, Belostotsky D, Bevan MW, Flavell RB, Rogers HJ, Lonsdale DM (1993) Partial characterization of the Nicotiana tabacum actin gene family: evidence for pollen specific expression of one of the gene family members. Mol Gen Genet 240:290–295
Twell D, Klein TM, Fromm ME, McCormick S (1989) Transient expression of chimeric genes delivered into pollen by microprojectile bombardment. Plant Physiol 91:1270–1274
Twell D, Yamaguchi J, McCormick S (1990) Pollen-specific gene expression in transgenic plants: coordinate regulation of two different tomato gene promoters during microsporogenesis. Genes Dev 109:705–713
Twell D, Yamaguchi J, Wing RA, Ushiba J, McCormick S (1991) Promoter analysis of genes that are coordinately expressed during pollen development reveals pollen-specific enhancer sequences and shared regulatory elements. Genes Dev 5: 496–507
van Tunen AJ, Hartman SA, Mur LA, Mol JNM (1989) Regulation of chalcone flavanone isomerase (CHI) gene expression in Petunia hybrida: the use of alternative promoters in corolla, anthers and pollen. Plant Mol Biol 12:539–551
Author information
Authors and Affiliations
Rights and permissions
About this article
Cite this article
Tebbutt, S.J., Lonsdale, D.M. Deletion analysis of a tobacco pollen-specific polygalacturonase promoter. Sexual Plant Reprod 8, 242–246 (1995). https://doi.org/10.1007/BF00228944
Received:
Accepted:
Issue Date:
DOI: https://doi.org/10.1007/BF00228944