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. 1991 Aug 25;19(16):4503–4508. doi: 10.1093/nar/19.16.4503

Stringent integrity requirements for both trans-activation and DNA-binding in a trans-activator, Oct3.

M Imagawa 1, A Miyamoto 1, M Shirakawa 1, H Hamada 1, M Muramatsu 1
PMCID: PMC328641  PMID: 1886774

Abstract

POU-specific and POU-homeo domains of Oct3 were produced in Echerichia coli for characterization of DNA binding to the octamer sequence. POU domain protein including A, B and H domains could bind to the octamer sequence efficiently and specifically, and DNase I footprint analysis gave an indistinguishable protection pattern between recombinant POU protein of Oct3 and native Oct3 from undifferentiated P19 cells. Truncated mutants, which contained B-specific and H domains or the H domain only, showed no binding activity, indicating that both of POU-specific and POU-homeo domains are essential for binding activity to octamer sequence. Furthermore, a 6 amino acid deletion from the N-terminal region of the A-specific domain is enough to destroy the binding activity. As for trans-activation, the N-terminal region is essential and sufficient. Deletion of the N-terminal proline-rich region rapidly eliminated trans-activating activity. These data strongly indicate the stringent integrity requirements for both trans-activation and DNA-binding domains in Oct3.

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