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Molecular and Cellular Biology, November 2007, p. 7981-7990, Vol. 27, No. 22
0270-7306/07/$08.00+0     doi:10.1128/MCB.01290-07
Copyright © 2007, American Society for Microbiology. All Rights Reserved.

p160 Myb-Binding Protein Interacts with Prep1 and Inhibits Its Transcriptional Activity{triangledown} ,{dagger}

Víctor M. Díaz,1,§ Silvia Mori,1 Elena Longobardi,1,4 Guillermo Menendez,1,{ddagger} Carmelo Ferrai,1 Rebecca A. Keough,2 Angela Bachi,3 and Francesco Blasi1,4*

Molecular Genetics Unit, Department of Molecular Biology and Functional Genomics, Università Vita Salute San Raffaele and DIBIT, H San Raffaele, via Olgettina 58, 20132 Milan, Italy,1 School of Molecular and Biomedical Sciences (Biochemistry), University of Adelaide, Adelaide, SA 5005, Australia,2 Mass Spectrometry Unit, San Raffaele Scientific Institute, via Olgettina 60, 20132 Milan, Italy,3 IFOM (FIRC Institute of Molecular Oncology), via Adamello 16, 20139 Milan, Italy4

Received 18 July 2007/ Returned for modification 21 August 2007/ Accepted 3 September 2007

Prep1 is known to interact in vivo with Pbx1 to regulate development and organogenesis. We have identified a novel Prep1-interacting protein, p160 c-Myb binding protein (p160). p160 and Pbx1 compete for Prep1 in vitro, and p160 inhibits Prep1-dependent HoxB2 expression in retinoic acid-treated NT2-D1 cells. The N-terminal physiologically truncated form of p160, p67, binds the sequence 63LFPLL67 in the HR1 domain of Prep1. Mutation of both L63 and L66 impairs the binding of Prep1 to both p160/p67 and Pbx1. The sequences required to bind Prep1 are mainly located in residues 51 to 151. Immunofluorescence colocalization and coimmunoprecipitation of endogenous p160 and Prep1 are induced by ActD, which translocates p160 from the nucleolus to the nucleoplasm. These data therefore show that p160 is a novel regulator of Prep1-Pbx1 transcriptional activity.

* Corresponding author. Mailing address: IFOM (FIRC Institute of Molecular Oncology), via Adamello 16, 20139 Milan, Italy. Phone: 0039 02 574303288. Fax: 0039 02 26434844. E-mail: blasi.francesco{at}hsr.it

{triangledown} Published ahead of print on 17 September 2007.

{dagger} Supplemental material for this article may be found at http://mcb.asm.org/.

§ Present address: Institut Municipal d'Investigació Biomèdica (IMIM), Parc de Recerca Biomèdica de Barcelona, Doctor Aiguader, 88, E-08003 Barcelona, Spain.

{ddagger} Present address: Molecular NeuroPathobiology Laboratory, Cancer Research UK, London Research Institute, 44 Lincoln's Inn Fields, London WC2A 3PX, United Kingdom.

Molecular and Cellular Biology, November 2007, p. 7981-7990, Vol. 27, No. 22
0270-7306/07/$08.00+0     doi:10.1128/MCB.01290-07
Copyright © 2007, American Society for Microbiology. All Rights Reserved.



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