Selective visualization of gene structure with ultraviolet light

Journal Article · Mon Feb 01 04:00:00 EST 1988 · Proc. Natl. Acad. Sci. U.S.A.; (United States)

DOI:https://doi.org/10.1073/pnas.85.3.654· OSTI ID:5314298

Wang, Z; Becker, M M

The ability of the ultraviolet (UV) footprinting technique to detect chromatin has been investigated in vitro. Two basic types of chromatin, a phased nucleosome and a phased nucleosome containing a phased H1 protein, have been reconstituted onto a cloned 5S ribosomal RNA gene from sea urchin. The histone-DNA interactions in each complex have been probed with exonuclease III, DNase I, dimethyl sulfate, and UV light. Whereas DNase I and exonuclease III readily detect interactions between histones and DNA, UV light and dimethyl sulfate do not. In contrast to histone-DNA interactions, we demonstrate that intimate sequence-specific contacts between the same sea urchin 5S DNA and the Xenopus laevis transcription factor IIIA (TFIIIA) are readily detected with UV light. Since the sensitivity of UV light for TFIIIA contacts is similar to its sensitivity for other regulatory protein-DNA contacts, these studies demonstrate the feasibility of using UV light to selectively visualize regulatory protein-DNA interactions in vivo with little or no interference from histone-DNA interactions.

• Research Organization: Univ. of Pittsburgh, PA (USA)
• OSTI ID: 5314298
• Journal Information: Proc. Natl. Acad. Sci. U.S.A.; (United States), Journal Name: Proc. Natl. Acad. Sci. U.S.A.; (United States) Vol. 85:3; ISSN PNASA
• Country of Publication: United States
• Language: English

Related Subjects

560120* -- Radiation Effects on Biochemicals
Cells
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63 RADIATION, THERMAL, AND OTHER ENVIRON. POLLUTANT EFFECTS ON LIVING ORGS. AND BIOL. MAT.
AMPHIBIANS
ANIMALS
AQUATIC ORGANISMS
CHROMATIN
DNA
DNA-ASE
ECHINODERMS
ELECTROMAGNETIC RADIATION
ENZYMES
ESTERASES
FROGS
GENES
GENETIC MAPPING
HISTONES
HYDROLASES
INVERTEBRATES
MAPPING
NUCLEASES
NUCLEIC ACIDS
NUCLEOPROTEINS
ORGANIC COMPOUNDS
OXYGEN COMPOUNDS
PHOSPHODIESTERASES
PROTEINS
RADIATIONS
RIBOSOMAL RNA
RNA
SEA URCHINS
SULFATES
SULFUR COMPOUNDS
TRANSCRIPTION
ULTRAVIOLET RADIATION
VERTEBRATES