NDLI: Functional Analysis of the 5′-Regulatory Region of the Maize ALKALINE ALPHA-GALACTOSIDASE1 Gene

Content Provider	Springer Nature Link
Author	Han, Qinghui Li, Tao Zhang, Lifeng Yan, Jun Dirk, Lynnette M.A. Downie, Bruce Zhao, Tianyong
Copyright Year	2014
Abstract	Maize ALKALINE ALPHA GALACTOSIDASE (ZmAGA) is a key enzyme in the raffinose metabolism pathway. We have previously characterized the ZmAGA1 gene as heat shock induced in germinating maize seeds. Here, we report that ZmAGA1 is induced in maize leaves by heat shock, dehydration, NaCl, abscisic acid (ABA), and salicylic acid. The 1622 bp of the 5′-regulatory region of the ZmAGA1 gene was isolated and characterized for promoter activity. The 5′-deletion assay of the regulatory region indicated that the region from −629 to −1 (relative to the start codon AUG) showed the greatest promoter activity, and the sequences upstream of this fragment act to inhibit transcription. The cis-acting element ABRE (ABA - responsive element) was identified to be functional. Increasingly, more refined 3′-deletion constructs of the regulatory region demonstrated that a 6-nucleotide (nt) RNA motif in the 5′-untranslated region (UTR) of the ZmAGA1 gene stimulates the translation of the downstream open reading frame (ORF). Deletion or triplication of this motif decreased or enhanced downstream ORF translation, respectively, while the mRNA abundance remained the same. RNA-EMSA (electrophoretic mobility shift assay) confirmed that some cytosolic protein(s) was (were) able to bind to this 6-nt RNA motif. This work contributes to our understanding of the ZmAGA1 gene regulatory mechanism and the metabolism of raffinose in maize cells.
Starting Page	1361
Ending Page	1370
Page Count	10
File Format	PDF
ISSN	07359640
Journal	Plant Molecular Biology Reporter
Volume Number	33
Issue Number	5
e-ISSN	15729818
Language	English
Publisher	Springer US
Publisher Date	2014-12-09
Publisher Place	New York
Access Restriction	One Nation One Subscription (ONOS)
Subject Keyword	Maize Raffinose Alkaline alpha-galactosidase Promoter Plant Sciences Plant Breeding/Biotechnology Proteomics Metabolomics Bioinformatics
Content Type	Text
Resource Type	Article
Subject	Plant Science Molecular Biology

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