DNA-binding factors assemble in a sequence-specific manner on the maize mitochondrial atpA promoter

Ching-Chun Chang, David B. Stern

Research output: Contribution to journalArticle

3 Citations (Scopus)

Abstract

The maize mitochondrial atpA promoter has been well-characterized using in vitro transcription. The functional elements of this promoter comprise a central domain extending from -7 to +5 relative to the transcription start site, and an upstream domain of 1-3 bp that is purine-rich and centered around positions -11 to -12. As a first step in characterizing the transcriptional machinery, exo-nuclease-III mapping (toeprinting) was used to map the borders of DNA-protein interactions using either a 107-bp wild-type template or transcriptionally-inactive templates containing linker-scanning mutations. These experiments revealed that, with a wild-type promoter, protein factors occupy as much as 36 bp, from positions -20 to +16 relative to the transcription initiation site. Protein-binding patterns were altered when the linker-scanning mutants were used, suggesting that either the number or conformation of DNA-binding proteins could account for their inability to promote transcription initiation.

Original languageEnglish
Pages (from-to)506-511
Number of pages6
JournalCurrent Genetics
Volume35
Issue number5
DOIs
Publication statusPublished - 1999 Jul 8

Fingerprint

Transcription Initiation Site
Zea mays
DNA
DNA-Binding Proteins
Protein Binding
Proteins
Mutation
purine
In Vitro Techniques

All Science Journal Classification (ASJC) codes

  • Genetics

Cite this

@article{2575a112cb1a464d84e9eb01cb674f6c,
title = "DNA-binding factors assemble in a sequence-specific manner on the maize mitochondrial atpA promoter",
abstract = "The maize mitochondrial atpA promoter has been well-characterized using in vitro transcription. The functional elements of this promoter comprise a central domain extending from -7 to +5 relative to the transcription start site, and an upstream domain of 1-3 bp that is purine-rich and centered around positions -11 to -12. As a first step in characterizing the transcriptional machinery, exo-nuclease-III mapping (toeprinting) was used to map the borders of DNA-protein interactions using either a 107-bp wild-type template or transcriptionally-inactive templates containing linker-scanning mutations. These experiments revealed that, with a wild-type promoter, protein factors occupy as much as 36 bp, from positions -20 to +16 relative to the transcription initiation site. Protein-binding patterns were altered when the linker-scanning mutants were used, suggesting that either the number or conformation of DNA-binding proteins could account for their inability to promote transcription initiation.",
author = "Ching-Chun Chang and Stern, {David B.}",
year = "1999",
month = "7",
day = "8",
doi = "10.1007/s002940050446",
language = "English",
volume = "35",
pages = "506--511",
journal = "Current Genetics",
issn = "0172-8083",
publisher = "Springer Verlag",
number = "5",

}

DNA-binding factors assemble in a sequence-specific manner on the maize mitochondrial atpA promoter. / Chang, Ching-Chun; Stern, David B.

In: Current Genetics, Vol. 35, No. 5, 08.07.1999, p. 506-511.

Research output: Contribution to journalArticle

TY - JOUR

T1 - DNA-binding factors assemble in a sequence-specific manner on the maize mitochondrial atpA promoter

AU - Chang, Ching-Chun

AU - Stern, David B.

PY - 1999/7/8

Y1 - 1999/7/8

N2 - The maize mitochondrial atpA promoter has been well-characterized using in vitro transcription. The functional elements of this promoter comprise a central domain extending from -7 to +5 relative to the transcription start site, and an upstream domain of 1-3 bp that is purine-rich and centered around positions -11 to -12. As a first step in characterizing the transcriptional machinery, exo-nuclease-III mapping (toeprinting) was used to map the borders of DNA-protein interactions using either a 107-bp wild-type template or transcriptionally-inactive templates containing linker-scanning mutations. These experiments revealed that, with a wild-type promoter, protein factors occupy as much as 36 bp, from positions -20 to +16 relative to the transcription initiation site. Protein-binding patterns were altered when the linker-scanning mutants were used, suggesting that either the number or conformation of DNA-binding proteins could account for their inability to promote transcription initiation.

AB - The maize mitochondrial atpA promoter has been well-characterized using in vitro transcription. The functional elements of this promoter comprise a central domain extending from -7 to +5 relative to the transcription start site, and an upstream domain of 1-3 bp that is purine-rich and centered around positions -11 to -12. As a first step in characterizing the transcriptional machinery, exo-nuclease-III mapping (toeprinting) was used to map the borders of DNA-protein interactions using either a 107-bp wild-type template or transcriptionally-inactive templates containing linker-scanning mutations. These experiments revealed that, with a wild-type promoter, protein factors occupy as much as 36 bp, from positions -20 to +16 relative to the transcription initiation site. Protein-binding patterns were altered when the linker-scanning mutants were used, suggesting that either the number or conformation of DNA-binding proteins could account for their inability to promote transcription initiation.

UR - http://www.scopus.com/inward/record.url?scp=0033066214&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=0033066214&partnerID=8YFLogxK

U2 - 10.1007/s002940050446

DO - 10.1007/s002940050446

M3 - Article

C2 - 10369957

AN - SCOPUS:0033066214

VL - 35

SP - 506

EP - 511

JO - Current Genetics

JF - Current Genetics

SN - 0172-8083

IS - 5

ER -