An 'environment to nucleus' signaling system operates in B lymphocytes: Redox status modulates BSAP/Pax-5 activation through Ref-1 nuclear translocation

Gianluca Tell, Alessandro Zecca, Lucia Pellizzari, Paola Spessotto, Alfonso Colombatti, Mark Kelley, Giuseppe Damante, Carlo Pucillo

Research output: Contribution to journalArticle

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Abstract

The Ref-1 (also called APE or HAP1) protein is a bifunctional enzyme impacting on a wide variety of important cellular functions. It acts as a major member of the DNA base excision repair pathway. Moreover, Ref-1 stimulates the DNA-binding activity of several transcription factors (TFs) through the reduction of highly reactive cysteine residues. Therefore, it represents a mechanism that regulates eukaryotic gene expression in a fast way. However, it has been demonstrated that external stimuli directly act on Ref-1 by increasing its expression levels, a time-consuming mechanism representing a paradox in terms of rapidity of TF regulation. In this paper we demonstrate that this is only an apparent paradox. Exposure of B lymphocytes to H2O2 induced a rapid and sustained increase in Ref-1 protein levels in the nucleus as evaluated by both western blot analysis and by pulse-chase experiments. A time course, two color in situ immunocytochemistry indicated that the up-regulation of Ref-1 in the nucleus at < 30 min was primarily the consequence of translocation of its cytoplasmic form. This early nuclear accumulation is effective in modulating the DNA-binding activity of the B cell-specific activator protein BSAP/Pax-5. In fact, EMSA experiments demonstrate that a transient interaction with Ref-1 up-regulates the DNA-binding activity of BSAP/Pax-5. Moreover, in a co-transfection experiment, Ref-1 increased the BSAP/Pax-5 activating effect on an oligomerized BSAP/Pax-5 binding site of the CD19 promoter by 5- to 8-fold. Thus, Ref-1 mediates its effect by up-regulating the DNA-binding activity of BSAP/Pax-5, accounting for a new and fast outside/inside pathway of signaling in B cells.

Original languageEnglish
Pages (from-to)1099-1105
Number of pages7
JournalNucleic Acids Research
Volume28
Issue number5
StatePublished - Mar 1 2000

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Oxidation-Reduction
B-Lymphocytes
DNA
PAX5 Transcription Factor
Transcription Factors
Up-Regulation
DNA Repair
Transfection
Cysteine
Proteins
Color
Western Blotting
Immunohistochemistry
Binding Sites
Gene Expression
Enzymes

ASJC Scopus subject areas

  • Genetics

Cite this

An 'environment to nucleus' signaling system operates in B lymphocytes : Redox status modulates BSAP/Pax-5 activation through Ref-1 nuclear translocation. / Tell, Gianluca; Zecca, Alessandro; Pellizzari, Lucia; Spessotto, Paola; Colombatti, Alfonso; Kelley, Mark; Damante, Giuseppe; Pucillo, Carlo.

In: Nucleic Acids Research, Vol. 28, No. 5, 01.03.2000, p. 1099-1105.

Research output: Contribution to journalArticle

Tell, G, Zecca, A, Pellizzari, L, Spessotto, P, Colombatti, A, Kelley, M, Damante, G & Pucillo, C 2000, 'An 'environment to nucleus' signaling system operates in B lymphocytes: Redox status modulates BSAP/Pax-5 activation through Ref-1 nuclear translocation', Nucleic Acids Research, vol. 28, no. 5, pp. 1099-1105.
Tell, Gianluca ; Zecca, Alessandro ; Pellizzari, Lucia ; Spessotto, Paola ; Colombatti, Alfonso ; Kelley, Mark ; Damante, Giuseppe ; Pucillo, Carlo. / An 'environment to nucleus' signaling system operates in B lymphocytes : Redox status modulates BSAP/Pax-5 activation through Ref-1 nuclear translocation. In: Nucleic Acids Research. 2000 ; Vol. 28, No. 5. pp. 1099-1105.
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