Online citations, reference lists, and bibliographies.
← Back to Search

Overproduction And Improved Strategies To Purify The Threenative Forms Of Nuclease-free HU Protein.

O. Pellegrini, J. Oberto, V. Pinson, M. Wery, J. Rouviere-Yaniv
Published 2000 · Medicine, Biology

Cite This
Download PDF
Analyze on Scholarcy
Share
The heterodimeric HU protein was isolated from Escherichia coli as one of the most abundant DNA binding proteins associated with the bacterial nucleoid. HUalphabeta is composed of two very homologous subunits, but HU can also be present in E. coli under its two homodimeric forms, HUalpha(2) and HUbeta(2). This protein is conserved either in its heterodimeric form or in one of its homodimeric forms in all bacteria, in plant chloroplasts and in some viruses. HU can participate, like the histones, in the maintenance of DNA supercoiling and in DNA condensation. This protein which does not recognize any specific sequence on double-stranded DNA, has been shown to bind specifically to cruciform DNA as does the eukaryotic HMG1 protein and to a series of structures which are found as intermediates of DNA repair, e.g., nick, gap, 3'overhang, etc. The strong binding of HU to these diverse DNA structures could explain, in part at least, its pleiotropic role in the bacterial cell. To understand all the facets of its interactions with nucleic acids, it was necessary to develop a procedure which allowed the purification of the three forms of HU under their native form and without the nuclease activity strongly associated with the protein. We describe here such a procedure as well as demonstrating that the three histidine-tagged HUs we have produced, have conserved the binding characteristics of native HUs. Interestingly, by two complementation tests, we show that the histidine-tagged HUs are fully active in vivo.
This paper references
10.1006/JMBI.1993.1656
An imbalance of HU synthesis induces mucoidy in Escherichia coli.
E. Painbéni (1993)
10.1016/0014-5793(84)80877-7
Similarity between the bacterial histone‐like protein HU and a protein from spinach chloroplasts
J. Briat (1984)
10.1006/JMBI.1999.2631
Differential binding of the Escherichia coli HU, homodimeric forms and heterodimeric form to linear, gapped and cruciform DNA.
V. Pinson (1999)
10.1016/0092-8674(91)90579-N
The phosphorylated form of the enhancer-binding protein NTRC has an ATPase activity that is essential for activation of transcription
D. Weiss (1991)
10.1016/0092-8674(79)90152-1
E. coli DNA binding protein HU forms nucleosome-like structure with circular double-stranded DNA
J. Rouvière-Yaniv (1979)
10.1128/JB.171.7.3704-3712.1989
Multiple defects in Escherichia coli mutants lacking HU protein.
O. Huisman (1989)
Molecular Cloning: A Laboratory Manual
J. Sambrook (1983)
10.1016/0014-5793(79)80518-9
Native Escherichia coli HU protein is a heterotypic dimer
J. Rouvière-Yaniv (1979)
10.1093/NAR/19.22.6337
Purification of his-tagged proteins in non-denaturing conditions suggests a convenient method for protein interaction studies.
A. Hoffmann (1991)
10.1006/JMBI.1994.1563
DNA-binding parameters of the HU protein of Escherichia coli to cruciform DNA.
E. Bonnefoy (1994)
10.1016/0022-2836(91)90855-Z
Use of bacteriophage T7 lysozyme to improve an inducible T7 expression system.
F. W. Studier (1991)
10.1111/j.1365-2958.1996.tb02476.x
The Escherichia coli ribosomal protein S16 is an endonuclease
J. Oberto (1996)
10.1073/pnas.74.12.5463
DNA sequencing with chain-terminating inhibitors.
F. Sanger (1977)
10.1073/PNAS.72.9.3428
Characterization of a novel, low-molecular-weight DNA-binding protein from Escherichia coli.
J. Rouvière-Yaniv (1975)
10.1006/JMBI.1997.1310
Variation in HU composition during growth of Escherichia coli: the heterodimer is required for long term survival.
L. Claret (1997)
10.1101/SQB.1978.042.01.047
Localization of the HU protein on the Escherichia coli nucleoid.
J. Rouviere-Yaniv (1978)
10.1128/JB.178.1.293-297.1996
Serratia marcescens contains a heterodimeric HU protein like Escherichia coli and Salmonella typhimurium.
J. Oberto (1996)
10.1073/PNAS.73.6.1917
Cyanobacterial DNA-binding protein related to Escherichia coli HU.
R. Haselkorn (1976)
10.1016/0300-9084(94)90014-0
Histones, HMG, HU, IHF: Même combat.
J. Oberto (1994)
10.1038/227680A0
Cleavage of Structural Proteins during the Assembly of the Head of Bacteriophage T4
U. Laemmli (1970)
10.1128/JB.173.9.2852-2863.1991
Inhibition of cell division in hupA hupB mutant bacteria lacking HU protein.
A. Dri (1991)
10.1128/JB.177.14.4121-4130.1995
Tight regulation, modulation, and high-level expression by vectors containing the arabinose PBAD promoter.
L. M. Guzmán (1995)



This paper is referenced by
10.1074/JBC.M400021200
Reconstitution of F Factor DNA Replication in Vitro with Purified Proteins*
S. Zzaman (2004)
10.1016/S0300-9084(01)01254-8
HU-GFP and DAPI co-localize on the Escherichia coli nucleoid.
M. Wery (2001)
10.1093/nar/gkm667
HU binds and folds single-stranded DNA
D. Kamashev (2008)
10.1007/s11008-005-0074-0
Comparison of the Local Concentration of the HU Protein for Particular Regions of Genomic DNA in Escherichia coli Cells in Vivo
O. V. Preobrajenskaya (2005)
10.1074/JBC.M308516200
Reconstitution of R6K DNA Replication in Vitro Using 22 Purified Proteins*
M. Abhyankar (2003)
10.1021/bi2009097
Mycoplasma gallisepticum produces a histone-like protein that recognizes base mismatches in DNA.
D. Kamashev (2011)
Etude structurale du mécanisme d'échange de chaînes des dimères de la protéine HU d'E. coli
R. L. Meur (2015)
10.1016/J.MOLCEL.2005.10.037
Oligomeric initiator protein-mediated DNA looping negatively regulates plasmid replication in vitro by preventing origin melting.
S. Zzaman (2005)
10.1371/journal.pone.0188037
Comparison of histone-like HU protein DNA-binding properties and HU/IHF protein sequence alignment
D. Kamashev (2017)
10.1046/j.1365-2958.2001.02305.x
The Escherichia coli histone‐like protein HU regulates rpoS translation
A. Balandina (2001)
10.1093/emboj/19.23.6527
The histone‐like protein HU binds specifically to DNA recombination and repair intermediates
D. Kamashev (2000)
10.1016/j.biochi.2011.03.005
Purification and functional analysis of recombinant Acholeplasma laidlawii histone-like HU protein.
S. Levitskiy (2011)
Semantic Scholar Logo Some data provided by SemanticScholar