DESIGN, SYNTHESIS, AND CHARACTERIZATION OF HIV-1 ENHANCER-BINDING POLYPEPTIDES DERIVED FROM BACTERIOPHAGE-434 REPRESSOR

作者：STADLER K^*; LIU NK; TROTMAN L; HILTPOLD A; CADERAS G; KLAUSER S; HEHLGANS T; GUTTE B

来源：International Journal of Peptide and Protein Research, 1995, 46(3-4): 333-340.

• ARTIFICIAL HIV ENHANCER-BINDING POLYPEPTIDES
• BACTERIOPHAGE-434 REPRESSOR-OPERATOR MODEL SYSTEM
• BAND SHIFT
• CIRCULAR DICHROISM
• DNASE I FOOT-PRINTING
• NF-KAPPA-B

We have designed and synthesized HIV-1 enhancer-binding polypeptides that were derived from bacterioph age 434 repressor. These peptides were 39-54 residues long and contained either the recognition helix or the entire helix-turn-helix motif of the DNA-binding domain of 434 repressor. The dissociation constant of the complex formed between the standard peptide (R42) and a synthetic 70-bp HIV enhancer DNA was ca. 10(-8) M. The specificity of the interaction of R42 with the two HIV enhancers was demonstrated by competitive band shift assays, stepwise displacement of the p50 subunit of transcription factor NF-kappa B from its two HIV enhancer binding sites, and DNase I footprinting; R42 seemed to protect best the two TTTCC sequences of the HIV enhancers against digestion by DNase I. R42 analogues with mutated recognition helix had lower DNA binding specificity. It remains to be investigated whether our artificial HIV enhancer-binding polypeptides are active in vivo.

• 出版日期1995-10