Artigo Revisado por pares

Structural Motif of the GCN4 DNA Binding Domain Characterized by Affinity Cleaving

1990; American Association for the Advancement of Science; Volume: 248; Issue: 4957 Linguagem: Inglês

10.1126/science.2111578

ISSN

1095-9203

Autores

Martha G. Oakley, Peter B. Dervan,

Tópico(s)

Protein Structure and Dynamics

Resumo

The NH2-terminal locations of a dimer containing the DNA binding domain of the yeast transcriptional activator GCN4 have been mapped on the binding sites 5'-CTGACTAAT-3' and 5'-ATGACTCTT-3'. Affinity cleaving was effected by synthetic GCN4 proteins with Fe.EDTA moieties at the NH2-terminus. Analysis of the DNA cleavage patterns for dimers of the Fe.EDTA-proteins corresponding to GCN4 residues 222 to 281 and 226 to 281 revealed that the NH2-termini were in the major groove nine to ten base pairs apart and were symmetrically displaced four to five base pairs from the central C of the recognition site. This result is consistent with the Y-shaped scissor grip-leucine zipper model recently proposed for a class of DNA binding proteins important in the regulation of gene expression.

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