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KMID : 0364219960390040393
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Korean Journal of Zoology
1996 Volume.39 No. 4 p.393 ~ p.399
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Zero-Length Crosslinking Study on Interactions of TBP, GAL4-AH, and TFIIB in the Preinitiation Complex
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Kwon Hyock-Man
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Abstract
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A zero-Iength croessinking procedure for studying protein-protein interactlons in preinitiation complex has heen developed. Preinltiadon complexes were assembled with immobilized DNA templates coupled to metal beads. Pudfied complexes were dIrectly crosslinked by 1-ethyl-3.(3-dimethylaminopropyl)carbodlimide (EDC). The reaction was stopped by addition of β-mercaptoethanol, and the complexes were isolated from EDC immedIately. An appllcatlion of this method with a preinltlation complex assembled with TBP, TFIIB, and GAL4-AH demonstrated that ThP dIrectly interacts with GAL4-AH and TFIIB in the prelnitlatlon complex. However, croeslinked produd between GAL4-AH and ThilB was not observed. These resutts lndlcate that GAL4-AH does not stably Interact with TFIIB In the GAL4-AH-TFIIB-TBP-DNA prelnitlatlon complex.
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KEYWORD
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TBP, GAL4-AH, TFIIB, Preinitiation Complex, Crosslinking
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