NDLI: Quantitative study of synthetic Hox transcription factor

Content Provider	PubMed Central
Author	Vukojević, Vladana Papadopoulos, Dimitrios K. Lars, Terenius Gehring, Walter J. Rigler, Rudolf
Abstract	Transcription factor–DNA interactions are life sustaining and therefore the subject of intensive research. In spite of vast effort, quantitative in vivo studies of the molecular mechanisms underlying these fundamental interactions remain challenging. In the preceding paper, we designed synthetic Sex combs reduced (Scr) peptides and validated genetically their function as transcriptional regulators. Here we present a controllable system for quantitative studies of protein–DNA interactions in live cells that enables us to “titrate” the concentration of the synthetic Scr peptides in a single cell. Using methods with single-molecule sensitivity, advanced fluorescence imaging and fluorescence correlation spectroscopy (FCS), we were able to study the kinetics of Scr-DNA interactions in live salivary gland cells, where Scr is normally expressed during development. We discerned freely moving Scr molecules, characterized the specific and nonspecific Scr peptide–DNA interactions, and estimated their corresponding dissociation constants (K d ) in vivo. Our results suggest that the synthetic Scr transcription factors find their specific target sites primarily by multiple association/dissociation events, the rapidity of which is largely owed to electrostatic interactions. Based on these new findings, we formulate a model mechanism and emulate the kinetics of Scr homeodomain–DNA interactions in live cells using numerical simulations.
Related Links	http://dx.doi.org/10.1073/pnas.0914612107
Ending Page	4098
Page Count	6
Starting Page	4093
File Format	PDF
ISSN	00278424
e-ISSN	10916490
Journal	Proceedings of the National Academy of Sciences of the United States of America
Issue Number	9
Volume Number	107
Language	English
Publisher	National Academy of Sciences
Publisher Date	2010-03-02
Access Restriction	Open
Rights Holder	National Academy of Sciences
Subject Keyword	General Research in Higher Education
Content Type	Text
Resource Type	Article
Subject	Multidisciplinary

Sl.	Authority	Responsibilities	Communication Details
1	Ministry of Education (GoI), Department of Higher Education	Sanctioning Authority	https://www.education.gov.in/ict-initiatives
2	Indian Institute of Technology Kharagpur	Host Institute of the Project: The host institute of the project is responsible for providing infrastructure support and hosting the project	https://www.iitkgp.ac.in
3	National Digital Library of India Office, Indian Institute of Technology Kharagpur	The administrative and infrastructural headquarters of the project	Dr. B. Sutradhar bsutra@ndl.gov.in
4	Project PI / Joint PI	Principal Investigator and Joint Principal Investigators of the project	Dr. B. Sutradhar bsutra@ndl.gov.in Prof. Saswat Chakrabarti will be added soon
5	Website/Portal (Helpdesk)	Queries regarding NDLI and its services	support@ndl.gov.in
6	Contents and Copyright Issues	Queries related to content curation and copyright issues	content@ndl.gov.in
7	National Digital Library of India Club (NDLI Club)	Queries related to NDLI Club formation, support, user awareness program, seminar/symposium, collaboration, social media, promotion, and outreach	clubsupport@ndl.gov.in
8	Digital Preservation Centre (DPC)	Assistance with digitizing and archiving copyright-free printed books	dpc@ndl.gov.in
9	IDR Setup or Support	Queries related to establishment and support of Institutional Digital Repository (IDR) and IDR workshops	idr@ndl.gov.in

Function and specificity of synthetic Hox transcription factors in vivo

Quantitative modeling of transcription factor binding specificities using DNA shape

Cross-regulatory protein–protein interactions between Hox and Pax transcription factors

INTERNAL REGULATORY INTERACTIONS DETERMINE DNA BINDING SPECIFICITY BY A HOX TRANSCRIPTION FACTOR

Surprising flexibility in a conserved Hox transcription factor over 550 million years of evolution

Labeling of fusion proteins with synthetic fluorophores in live cells

Systematic measurement of transcription factor-DNA interactions by targeted mass spectrometry identifies candidate gene regulatory proteins

Role for β-catenin and HOX transcription factors in Caenorhabditis elegans and mammalian host epithelial-pathogen interactions

A Live Cell Protein Complementation Assay for ORFeome-Wide Probing of Human HOX Interactomes

Quantitative study of synthetic Hox transcription factor–DNA interactions in live cells

Similar Documents

Function and specificity of synthetic Hox transcription factors in vivo

Quantitative modeling of transcription factor binding specificities using DNA shape

Cross-regulatory protein–protein interactions between Hox and Pax transcription factors

INTERNAL REGULATORY INTERACTIONS DETERMINE DNA BINDING SPECIFICITY BY A HOX TRANSCRIPTION FACTOR

Surprising flexibility in a conserved Hox transcription factor over 550 million years of evolution

Labeling of fusion proteins with synthetic fluorophores in live cells

Systematic measurement of transcription factor-DNA interactions by targeted mass spectrometry identifies candidate gene regulatory proteins

Role for β-catenin and HOX transcription factors in Caenorhabditis elegans and mammalian host epithelial-pathogen interactions

A Live Cell Protein Complementation Assay for ORFeome-Wide Probing of Human HOX Interactomes

Quantitative study of synthetic Hox transcription factor–DNA interactions in live cells