Co-crystallization of an ETS domain (PU.1) in complex with DNA: Engineering the length of both protein and oligonucleotide

Frédéric Pio, Chao Zhou Ni, Richard S. Mitchell, John Knight, Scott McKercher, Michael Klemsz, Angela Lombardo, Richard A. Maki, Kathryn R. Ely

Research output: Contribution to journalArticle

14 Citations (Scopus)

Abstract

The PU.1 transcription factor is a member of the ets gene family of regulatory proteins. These molecules play a role in normal development and also have been implicated in malignant processes such as the development of erythroid leukemia. The Ets proteins share a conserved DNA-binding domain (the ETS domain) that recognizes a purine-rich sequence with the core sequence: 5′-Cl AGGAA/T-3′. This domain binds to DNA as a monomer, unlike many other DNA-binding proteins. The ETS domain of the PU.1 transcription factor has been crystallized in complex with a 16-base pair oligonucleotide that contains the recognition sequence. The crystals formed in the space group C2 with a = 89.1, b = 101.9, c = 55.6 Å, and β= 111.2° and diffract to at least 2.3 Å. There are two complexes in the asymmetric unit. Production of large usable crystals was dependent on the length of both protein and DNA components, the use of oligonucleotides with unpaired A and T bases at the termini, and the presence of polyethylene glycol and zinc acetate in the crystallization solutions. This is the first ETS domain to be crystallized, and the strategy used to crystallize this complex may be useful for other members of the ets family.

Original languageEnglish
Pages (from-to)24258-24263
Number of pages6
JournalJournal of Biological Chemistry
Volume270
Issue number41
StatePublished - Oct 13 1995

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Crystallization
Oligonucleotides
DNA
Transcription Factors
Zinc Acetate
Crystals
Proteins
DNA-Binding Proteins
Regulator Genes
Base Pairing
Leukemia
Genes
Monomers
Molecules
ETS Motif

ASJC Scopus subject areas

  • Biochemistry

Cite this

Pio, F., Ni, C. Z., Mitchell, R. S., Knight, J., McKercher, S., Klemsz, M., ... Ely, K. R. (1995). Co-crystallization of an ETS domain (PU.1) in complex with DNA: Engineering the length of both protein and oligonucleotide. Journal of Biological Chemistry, 270(41), 24258-24263.

Co-crystallization of an ETS domain (PU.1) in complex with DNA : Engineering the length of both protein and oligonucleotide. / Pio, Frédéric; Ni, Chao Zhou; Mitchell, Richard S.; Knight, John; McKercher, Scott; Klemsz, Michael; Lombardo, Angela; Maki, Richard A.; Ely, Kathryn R.

In: Journal of Biological Chemistry, Vol. 270, No. 41, 13.10.1995, p. 24258-24263.

Research output: Contribution to journalArticle

Pio, F, Ni, CZ, Mitchell, RS, Knight, J, McKercher, S, Klemsz, M, Lombardo, A, Maki, RA & Ely, KR 1995, 'Co-crystallization of an ETS domain (PU.1) in complex with DNA: Engineering the length of both protein and oligonucleotide', Journal of Biological Chemistry, vol. 270, no. 41, pp. 24258-24263.
Pio, Frédéric ; Ni, Chao Zhou ; Mitchell, Richard S. ; Knight, John ; McKercher, Scott ; Klemsz, Michael ; Lombardo, Angela ; Maki, Richard A. ; Ely, Kathryn R. / Co-crystallization of an ETS domain (PU.1) in complex with DNA : Engineering the length of both protein and oligonucleotide. In: Journal of Biological Chemistry. 1995 ; Vol. 270, No. 41. pp. 24258-24263.
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