Tissue-specific expression of the PNZIP promoter is mediated by combinatorial interaction of different cis-elements and a novel transcriptional factor.
Recent studies demonstrated that PNZIP and its homologs encode a special cyclase and play an important role in chlorophyll biosynthesis in higher plants. To investigate the molecular mechanism governing the PNZIP gene, the PNZIP promoter was isolated and analyzed. Deletion analysis indicated that G-box is an important element in the regulation of the reporter gene expression. Further mutation assay demonstrated that G-box and GATACT elements are necessary and sufficient for the high and tissue-specific expression of the GUS gene. Using yeast one-hybrid screening, we have isolated a novel tobacco bZIP protein, NtbZIP, which can specifically recognize the G-box of the PNZIP promoter. The NtbZIP protein shares a limited amino acid homology to Arabidopsis ABI5 and AtAREB1 and very low homology to other bZIP proteins. Northern blot analysis showed that the NtbZIP gene is not induced by exogenous ABA and is expressed in different tobacco organs. Cotransformation assays showed that the NtbZIP protein could activate the transcription of the GUS gene driven by the PNZIP promoter. Transgenic tobaccos analysis demonstrated that constitutively expressing antisense NtbZIP gene resulted in a lower NTZIP synthesis and reduced chlorophyll levels. We suggest that NTZIP is a target gene of NtbZIP, which is involved in the regulation of chlorophyll biosynthesis.
Yang
Yu-Tao
YT
State Key Laboratory of Crop Biology, College of Life Sciences, Shandong Agricultural University, Taian, Shandong 271018, China.
Yu
Yan-Li
YL
Yang
Guo-Dong
GD
Zhang
Jie-Dao
JD
Zheng
Cheng-Chao
CC
D000595:Amino Acid Sequence; D001483:Base Sequence; D001665:Binding Sites; D018076:DNA, Complementary / Q000302:isolation & purification; D004268:DNA-Binding Proteins / Q000235:genetics; D018506:Gene Expression Regulation, Plant; D017930:Genes, Reporter; D008969:Molecular Sequence Data; D009687:Nuclear Proteins / Q000032:analysis; D010105:Oxygenases / Q000096:biosynthesis / Q000235:genetics*; D010788:Photosynthesis / Q000235:genetics; D010940:Plant Proteins / Q000096:biosynthesis / Q000235:genetics* / Q000378:metabolism; D011401:Promoter Regions, Genetic; D020218:Response Elements; D017421:Sequence Analysis; D014018:Tissue Distribution; D014026:Nicotiana / Q000201:enzymology / Q000235:genetics* / Q000378:metabolism; D015534:Trans-Activators / Q000032:analysis / Q000378:metabolism*
D018076:DNA, Complementary; D004268:DNA-Binding Proteins; D009687:Nuclear Proteins; D010940:Plant Proteins; D015534:Trans-Activators; D010105:Oxygenases; C053161:magnesium protoporphyrin monomethyl ester oxidative cyclase