Specific transcription and reinitiation of 2,4-D-induced genes in tobacco nuclei

• Published in: Plant molecular biology Vol. 18; no. 1; pp. 109 - 117
• Main Authors: Mennes, A.M. (Leiden Univ. (Netherlands). Dept. of Plant Molecular Biology, Botanical Lab.), Quint, A, Gribnau, J.H, Boot, C.J.M, Zaal, E.J. van der, Maan, A.C, Libbenga, K.R
• Format: Journal Article
• Language: English
• Published: Dordrecht Springer 1992
• Subjects: 2,4-D; 2,4-Dichlorophenoxyacetic Acid - pharmacology; ARN; AUXINAS; AUXINE; AUXINS; Base Sequence; Biological and medical sciences; Blotting, Northern; Cell Nucleus - metabolism; Chromatography, Affinity; EXPERIMENTACION IN VITRO; EXPERIMENTATION IN VITRO; Fundamental and applied biological sciences. Psychology; GENE; Gene Expression Regulation - drug effects; GENES; HEPARIN; Heparin - pharmacology; HEPARINA; HEPARINE; IN VITRO EXPERIMENTATION; Indoleacetic Acids - pharmacology; Molecular and cellular biology; Molecular genetics; Molecular Sequence Data; NICOTIANA; Nicotiana - genetics; NOYAU CELLULAIRE; NUCLEO; NUCLEUS; Plants, Toxic; RNA; Transcription, Genetic - genetics; Transcription. Transcription factor. Splicing. Rna processing; In vitro transcription; Auxin; Stimulant plant; Nicotiana tabacum; In vitro; Messenger RNA; Gene; Dicotyledones; Angiospermae; Plant growth substance; Trans acting regulatory factor; Spermatophyta; Solanaceae; Transcription initiation
• ISSN: 0167-4412; 1573-5028
• DOI: 10.1007/BF00018461

In order to study the effects of trans-acting factors on cis-acting elements in plant genes an in vitro reinitiating nuclear transcription system is needed. Here we report that run-on experiments with nuclei isolated from 2,4-D-treated auxin-starved early-stationary-phase cells of tobacco clearly show reitintiation of transcription of specific 2,4-D-induced genes. Using gamma-thio nucleotides and [alpha-32P]-UTP we were able to demonstrate the presence of reinitiated labelled specific RNAs after isolation on a mercury-agarose affinity column. Addition of heparin as an inhibitor blocked this reinitiation. In a primer extension assay we found that the new transcripts initiate at approximately the same site as used in vivo.