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PMID: 9171102 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't

Crosslinking of progesterone receptor to DNA using tuneable nanosecond, picosecond and femtosecond UV laser pulses.

Nucleic acids research ·Vol. 25 ·No. 12 ·1997-06-15 ·Pages 2478-84

Russmann C, Truss M, Fix A, Naumer C, Herrmann T, Schmitt J, Stollhof J, Beigang R, Beato M

Abstract

UV laser crosslinking is a potentially powerful tool to investigate transient DNA-protein interactions and binding kinetics in intact cells. As the processes underlying UV laser crosslinking are not fully understood, we have performed a study of the influence of laser pulses with different physical parameters on crosslinking of the progesterone receptor to an oligonucleotide containing a hormone-responsive element. We also studied the influence of the various parameters on the amount of laser-irradiated DNA that can be correctly primer extended as an operational measurement of DNA integrity. A strong influence of pulse intensity and pulse length on the crosslink yield was found, likely due to a change in the 'two photon' processes responsible for crosslinking. The highest efficiency of protein crosslinking to DNA was achieved with femtosecond pulses and should be sufficient to enable use of this technique for in vivo studies.

MeSH Terms

Animals Cross-Linking Reagents DNA Damage DNA, Viral/isolation & purification,metabolism,radiation effects Kinetics Lasers Mammary Tumor Virus, Mouse/genetics Promoter Regions, Genetic Rabbits Receptors, Progesterone/isolation & purification,metabolism,radiation effects Recombinant Proteins/metabolism,radiation effects Time Factors Ultraviolet Rays

Chemicals

Cross-Linking Reagents DNA, Viral Receptors, Progesterone Recombinant Proteins

Authors & Affiliations

9 authors, click to expand affiliations / ORCID

Russmann C

Institut für Molekularbiologie und Tumorforschung, Emil-Mannkopff-Strasse 2, D-35033 Marburg, Germany.

Truss M

Fix A

Naumer C

Herrmann T

Schmitt J

Stollhof J

Beigang R

Beato M

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Article Info

Journal

Nucleic acids research

Abbr.

Nucleic Acids Res

ISSN

0305-1048

DOI

10.1093/nar/25.12.2478

Published

1997-06-15

Pages

2478-84

Language

English

Region

England

NLM ID

0411011

PMCID

PMC146765

Subset

External Links

PubMed source DOI: 10.1093/nar/25.12.2478 PMC Full Text View journal details

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Crosslinking of progesterone receptor to DNA using tuneable nanosecond, picosecond and femtosecond UV laser pulses.

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