NMR study of the interaction of the HU protein from Bacillus stearothermophilus with DNA

Hans Vis; Olga Vageli; Jord Nagel; Constantin E. Vorgias; Keith S. Wilson; Robert Kaptein; Rolf Boelens

doi:10.1002/(sici)1097-458x(199612)34:13<s81::aid-omr67>3.0.co;2-7

NMR study of the interaction of the HU protein from Bacillus stearothermophilus with DNA

Hans Vis, Olga Vageli, Jord Nagel, Constantin E. Vorgias, Keith S. Wilson, Robert Kaptein^*, Rolf Boelens

^*Corresponding author for this work

Research output: Contribution to journal › Article › Academic › peer-review

Abstract

DNA binding of the 19.5 kDa protein HU from Bacillus stearothermophilus (HUBst) with several non-specific double-stranded DNA oligomers was studied using NMR techniques. Photochemically induced nuclear polarization (photo-CIDNP) measurements on a mutant HUBst (M69Y; V76Y) show that Y69 in the tip of the β-hairpin arm is involved in DNA interaction. Changes in ¹⁵N and ¹H^N chemical shifts upon titrating DNA are greatest for the residues in the β-hairpin arm and for the residues in the second half of the third a-helix. The changes in the flexible arms can be interpreted as being due to the formation of more rigid secondary structure upon DNA binding. Backbone and side-chain dynamics of HUBst were investigated using heteronuclear ¹⁵N-¹H^N NOE, ¹⁵N T₁ and T_1p data for free and complexed HUBst. These measurements show that the mobility of the flexible arms reduces in the protein-DNA complex. The results demonstrate that the β-hairpin arms and the C-terminal α-helix of HUBst are involved in DNA binding.

Original language	English
Pages (from-to)	S81-S86
Journal	Magnetic Resonance in Chemistry
Volume	34
Issue number	999
DOIs	https://doi.org/10.1002/(sici)1097-458x(199612)34:13<s81::aid-omr67>3.0.co;2-7
Publication status	Published - Dec 1996

Keywords

N NMR
H NMR
DNA-binding protein
Heteronuclear NOE
Mutant HU
NMR
Photo-CIDNP
Relaxation measurements

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10.1002/(sici)1097-458x(199612)34:13<s81::aid-omr67>3.0.co;2-7

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title = "NMR study of the interaction of the HU protein from Bacillus stearothermophilus with DNA",

abstract = "DNA binding of the 19.5 kDa protein HU from Bacillus stearothermophilus (HUBst) with several non-specific double-stranded DNA oligomers was studied using NMR techniques. Photochemically induced nuclear polarization (photo-CIDNP) measurements on a mutant HUBst (M69Y; V76Y) show that Y69 in the tip of the β-hairpin arm is involved in DNA interaction. Changes in 15N and 1HN chemical shifts upon titrating DNA are greatest for the residues in the β-hairpin arm and for the residues in the second half of the third a-helix. The changes in the flexible arms can be interpreted as being due to the formation of more rigid secondary structure upon DNA binding. Backbone and side-chain dynamics of HUBst were investigated using heteronuclear 15N-1HN NOE, 15N T1 and T1p data for free and complexed HUBst. These measurements show that the mobility of the flexible arms reduces in the protein-DNA complex. The results demonstrate that the β-hairpin arms and the C-terminal α-helix of HUBst are involved in DNA binding.",

keywords = "N NMR, H NMR, DNA-binding protein, Heteronuclear NOE, Mutant HU, NMR, Photo-CIDNP, Relaxation measurements",

author = "Hans Vis and Olga Vageli and Jord Nagel and Vorgias, {Constantin E.} and Wilson, {Keith S.} and Robert Kaptein and Rolf Boelens",

year = "1996",

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doi = "10.1002/(sici)1097-458x(199612)34:13<s81::aid-omr67>3.0.co;2-7",

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T1 - NMR study of the interaction of the HU protein from Bacillus stearothermophilus with DNA

AU - Vis, Hans

AU - Vageli, Olga

AU - Nagel, Jord

AU - Vorgias, Constantin E.

AU - Wilson, Keith S.

AU - Kaptein, Robert

AU - Boelens, Rolf

PY - 1996/12

Y1 - 1996/12

N2 - DNA binding of the 19.5 kDa protein HU from Bacillus stearothermophilus (HUBst) with several non-specific double-stranded DNA oligomers was studied using NMR techniques. Photochemically induced nuclear polarization (photo-CIDNP) measurements on a mutant HUBst (M69Y; V76Y) show that Y69 in the tip of the β-hairpin arm is involved in DNA interaction. Changes in 15N and 1HN chemical shifts upon titrating DNA are greatest for the residues in the β-hairpin arm and for the residues in the second half of the third a-helix. The changes in the flexible arms can be interpreted as being due to the formation of more rigid secondary structure upon DNA binding. Backbone and side-chain dynamics of HUBst were investigated using heteronuclear 15N-1HN NOE, 15N T1 and T1p data for free and complexed HUBst. These measurements show that the mobility of the flexible arms reduces in the protein-DNA complex. The results demonstrate that the β-hairpin arms and the C-terminal α-helix of HUBst are involved in DNA binding.

AB - DNA binding of the 19.5 kDa protein HU from Bacillus stearothermophilus (HUBst) with several non-specific double-stranded DNA oligomers was studied using NMR techniques. Photochemically induced nuclear polarization (photo-CIDNP) measurements on a mutant HUBst (M69Y; V76Y) show that Y69 in the tip of the β-hairpin arm is involved in DNA interaction. Changes in 15N and 1HN chemical shifts upon titrating DNA are greatest for the residues in the β-hairpin arm and for the residues in the second half of the third a-helix. The changes in the flexible arms can be interpreted as being due to the formation of more rigid secondary structure upon DNA binding. Backbone and side-chain dynamics of HUBst were investigated using heteronuclear 15N-1HN NOE, 15N T1 and T1p data for free and complexed HUBst. These measurements show that the mobility of the flexible arms reduces in the protein-DNA complex. The results demonstrate that the β-hairpin arms and the C-terminal α-helix of HUBst are involved in DNA binding.

KW - N NMR

KW - H NMR

KW - DNA-binding protein

KW - Heteronuclear NOE

KW - Mutant HU

KW - NMR

KW - Photo-CIDNP

KW - Relaxation measurements

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SP - S81-S86

JO - Magnetic Resonance in Chemistry

JF - Magnetic Resonance in Chemistry

IS - 999

ER -

NMR study of the interaction of the HU protein from Bacillus stearothermophilus with DNA

Abstract

Keywords

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