Jove
Visualize
Contact Us
JoVE
x logofacebook logolinkedin logoyoutube logo
ABOUT JoVE
OverviewLeadershipBlogJoVE Help Center
AUTHORS
Publishing ProcessEditorial BoardScope & PoliciesPeer ReviewFAQSubmit
LIBRARIANS
TestimonialsSubscriptionsAccessResourcesLibrary Advisory BoardFAQ
RESEARCH
JoVE JournalMethods CollectionsJoVE Encyclopedia of ExperimentsArchive
EDUCATION
JoVE CoreJoVE BusinessJoVE Science EducationJoVE Lab ManualFaculty Resource CenterFaculty Site
Terms & Conditions of Use
Privacy Policy
Policies

Related Experiment Videos

Initial transcribed sequence mutations specifically affect promoter escape properties.

Lilian M Hsu1, Ingrid M Cobb, Jillian R Ozmore

  • 1Program in Biochemistry, Mount Holyoke College, South Hadley, Massachusetts 01075, USA. lhsu@mtholyoke.edu

Biochemistry
|July 19, 2006
PubMed
Summary
This summary is machine-generated.

Related Concept Videos

You might also read

Related Articles

Articles linked to this work by shared authors, journal, and citation graph.

Sort by
Same author

Quantitative parameters of productive transcription on T5 N25-based promoters are modulated by the initial transcribed sequence and template supercoiling.

The Journal of biological chemistry·2025
Same author

SEMA6B variants cause intellectual disability and alter dendritic spine density and axon guidance.

Human molecular genetics·2022
Same author

A Novel SPAST/SPG4 Splice-Site Variant in a Family with Dominant Hereditary Spastic Paraplegia.

Case reports in neurological medicine·2020
Same author

Mechanisms of Very Long Abortive Transcript Release during Promoter Escape.

Biochemistry·2015
Same author

Promoter Escape by Escherichia coli RNA Polymerase.

EcoSal Plus·2015
Same author

Sequence-Dependent Promoter Escape Efficiency Is Strongly Influenced by Bias for the Pretranslocated State during Initial Transcription.

Biochemistry·2015
Same journal

Aromatic Cage-Directed Azide-Methyllysine Photochemistry for Profiling Nonhistone Interacting Partners of the MeCP2 Methyl-CpG-Binding Domain.

Biochemistry·2026
Same journal

Differential Hydroxypyruvate Processing by <i>E. coli</i> and <i>P. aeruginosa</i> DXP Synthases Reveals Preferential Xylulose 5-Phosphate Formation by the <i>P. aeruginosa</i> Enzyme.

Biochemistry·2026
Same journal

Structural and Functional Characterization of Heterologous Nitrogenase Complexes.

Biochemistry·2026
Same journal

Discovery of Bacterial Unspecific Peroxygenases.

Biochemistry·2026
Same journal

Lactate Biology: Subcellular Routing and Chemical Form Define Function.

Biochemistry·2026
Same journal

Nature's Anaerobic Toolkit: Glycyl Radical Enzymes and Their Expanding Functional and Mechanistic Diversity.

Biochemistry·2026
See all related articles

The initial transcribed sequence (ITS) significantly impacts bacterial RNA polymerase promoter escape efficiency. Variants of the T5 N25 promoter showed altered escape sites and efficiency, with the natural promoter optimized for rapid escape.

Area of Science:

  • Molecular Biology
  • Genetics
  • Biochemistry

Background:

  • Promoter escape is a critical step in bacterial transcription initiation.
  • The core promoter and initial transcribed sequence (ITS) influence RNA polymerase promoter escape efficiency.

Purpose of the Study:

  • To quantitatively examine the promoter escape properties of E. coli RNA polymerase using variants of the phage T5 N25 promoter's initial transcribed sequence (ITS).
  • To understand the role of ITS composition and the GreB factor in abortive initiation and promoter escape.

Main Methods:

  • Generated and analyzed 43 random initial sequence variants of the phage T5 N25 promoter.
  • Supplemented GreB factor into transcription reactions to study abortive initiation and promoter escape in vivo.
  • Analyzed abortive RNA patterns to assess the impact of GreB.

Related Experiment Videos

Main Results:

  • Promoter escape for N25-ITS variants occurred at the +15/+16 juncture, differing from the wild type's +11/+12 juncture.
  • Variants showed a 25-fold difference in escape efficiency, influenced by ITS composition unrelated to nucleotide binding.
  • GreB supplementation increased productive RNA synthesis 2-5 fold by altering abortive RNA profiles, decreasing medium and long abortive RNAs.

Conclusions:

  • The natural N25 promoter's ITS is optimized for efficient promoter escape, minimizing abortive initiation.
  • Short abortive RNAs may require a minimum RNA-DNA hybrid length for complex stability.
  • GreB's effect on abortive RNAs provides insights into polymerase backtracking and initial transcribing complex dynamics.