Distinct classes of yeast promoters revealed by differential TAF recruitment

Science

Volumn 288, Issue 5469, 2000, Pages 1242-1244

  Li, Xiao Yong ^a   Bhaumik, Sukesh R ^a   Green, Michael R ^a  

^a NONE

Author keywords

[No Author keywords available]

Indexed keywords

TATA BINDING PROTEIN; TRANSCRIPTION FACTOR;

ARTICLE; GENE FUNCTION; GENE INACTIVATION; GENE TARGETING; GENETIC ASSOCIATION; GENETIC TRANSCRIPTION; NONHUMAN; PRIORITY JOURNAL; PROMOTER REGION; YEAST;

CROSS-LINKING REAGENTS; DNA; DNA-BINDING PROTEINS; FORMALDEHYDE; MUTATION; PRECIPITIN TESTS; PROMOTER REGIONS (GENETICS); SACCHAROMYCES CEREVISIAE; TATA BOX; TATA-BOX BINDING PROTEIN; TRANS-ACTIVATORS; TRANSCRIPTION FACTORS; TRANSCRIPTION FACTORS, TFII;

EID: 0034686001     PISSN: 00368075     EISSN: None     Source Type: Journal    
DOI: 10.1126/science.288.5469.1242     Document Type: Article

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10. 2+) genes also was not affected after inactivation of any of the other known yeast TAFs (data not shown).
11. In brief, the crosslinking procedure is as follows. Live yeast cells were treated with 1% formaldehyde to induce crosslinks and disrupted with glass beads; the resultant extract was sonicated to generate DNA fragments with an average size of about 500 base pairs. Crosslinked complexes were immunoprecipitated with the specified antibodies, and proteinase K was used to release immunoprecipitated DNA from protein A-agarose beads. DNA samples were incubated at 65°C for 6 hours to reverse the crosslinks, purified, and used for quantitative polymerase chain reaction (PCR) analysis. Under these conditions, the crosslinking assay provides a quantitative measure of factor association with the promoter (11).
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25. Supported in part by NIH grant GM33977. We thank S. Buratowski, K. Struhl, and R. Young for providing yeast strains and K. Struhl for communicating results before publication.