Specific transcription and reinitiation of 2,4-D-induced genes in tobacco nuclei.
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.
Mennes
A M
AM
Department of Plant Molecular Biology, Leiden University, Netherlands.
Quint
A
A
Gribnau
J H
JH
Boot
C J
CJ
van der Zaal
E J
EJ
Maan
A C
AC
Libbenga
K R
KR
D015084:2,4-Dichlorophenoxyacetic Acid / Q000494:pharmacology*; D001483:Base Sequence; D015152:Blotting, Northern; D002467:Cell Nucleus / Q000378:metabolism; D002846:Chromatography, Affinity; D005786:Gene Expression Regulation / Q000187:drug effects*; D006493:Heparin / Q000494:pharmacology; D007210:Indoleacetic Acids / Q000494:pharmacology*; D008969:Molecular Sequence Data; D010947:Plants, Toxic; D014026:Nicotiana / Q000235:genetics*; D014158:Transcription, Genetic / Q000235:genetics*
D007210:Indoleacetic Acids; D015084:2,4-Dichlorophenoxyacetic Acid; D006493:Heparin