Exchange of histidine spacing between Sp1 and GLI zinc fingers: distinct effect of histidine spacing-linker region on DNA binding.

Abstract:

:In the DNA recognition mode of C(2)H(2)-type zinc fingers, the finger-finger connection region, consisting of the histidine spacing (HX(3-5)H) and linker, would be important for determining the orientation of the zinc finger domains. To clarify the influence of spacing between two ligand histidines in the DNA binding, we exchanged the histidine spacing between Sp1 and GLI zinc fingers, which have an HX(3)H-TGEKK linker (typical) and an HX(4)H-SNEKP linker (atypical), respectively. A significant decrease in the DNA binding affinity and specificity is found in Sp1-type peptides, whereas GLI-type peptides show a mild reduction. To evaluate the effect of the linker characteristics, we further designed Sp1-type mutants with an SNEKP linker. As a result, the significant effect of the histidine spacing in Sp1-type peptides was reduced. These results demonstrate that (1) the histidine spacing significantly affects the DNA binding of zinc finger proteins and (2) the histidine spacing and the following linker regions are one effective target for regulating the DNA recognition mode of zinc finger proteins.

journal_name

Biochemistry

journal_title

Biochemistry

authors

Shiraishi Y,Imanishi M,Sugiura Y

doi

10.1021/bi035999s

subject

Has Abstract

pub_date

2004-05-25 00:00:00

pages

6352-9

issue

20

eissn

0006-2960

issn

1520-4995

journal_volume

43

pub_type

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