Purification of the Prep1 interactome identifies novel pathways regulated by Prep1.
Prep1 homeodomain transcription factor interacts with Pbx proteins to regulate oculogenesis, angiogenesis, and hematopoiesis in mice. To isolate new Prep1 interactors competing or copurifying with Pbx, we identified proteins copurified with Prep1-TAP by tandem affinity purification (TAP). Prep1-TAP was fully functional and allowed the isolation of a Prep1 proteome from cytoplasm and nucleus, but most interactors were nuclear. The Prep1-TAP complex included Pbx1b, Pbx2, and other nonhomeodomain proteins: p160 Myb-binding protein (p160), beta-actin, NMMHCIIA.
Díaz
Víctor M
VM
Molecular Genetics Unit, Department of Molecular Biology and Functional Genomics, Università Vita Salute San Raffaele and DIBIT, H San Raffaele, Milan, Italy.
Bachi
Angela
A
Blasi
Francesco
F
D000818:Animals; D006690:Bisbenzimidazole / Q000378:metabolism; D002467:Cell Nucleus / Q000378:metabolism; D002846:Chromatography, Affinity; D019084:Fluorescent Antibody Technique, Indirect; D005456:Fluorescent Dyes / Q000378:metabolism; D005786:Gene Expression Regulation / Q000502:physiology*; D017930:Genes, Reporter; D005822:Genetic Vectors; D018398:Homeodomain Proteins / Q000737:chemistry / Q000302:isolation & purification* / Q000378:metabolism* / Q000494:pharmacology; D008156:Luciferases / Q000378:metabolism; D013058:Mass Spectrometry; D051379:Mice; D041681:NIH 3T3 Cells; D011485:Protein Binding; D017434:Protein Structure, Tertiary; D011500:Protein Synthesis Inhibitors / Q000494:pharmacology; D011691:Puromycin / Q000494:pharmacology; D012190:Retroviridae / Q000235:genetics; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization; D014157:Transcription Factors / Q000737:chemistry / Q000302:isolation & purification* / Q000378:metabolism* / Q000494:pharmacology; D014162:Transfection
D005456:Fluorescent Dyes; D018398:Homeodomain Proteins; C491412:Pknox1 protein, mouse; D011500:Protein Synthesis Inhibitors; D014157:Transcription Factors; D011691:Puromycin; D008156:Luciferases; D006690:Bisbenzimidazole