title: PCNA interacts with Prox1 and represses its transcriptional activity.
abstract: OBJECTIVE
  Prox1 is a transcription factor which can function either as a transcriptional activator, transcriptional repressor or a transcriptional corepressor. This paper seeks to better understand the role of protein-protein interactions in this multitude of functions.
  
  
  METHODS
  We performed a yeast two-hybrid screen of an 11.5 day post coitum (dpc) mouse embryo cDNA library using the homeo-Prospero domain of Prox1 as bait. Computer modeling, cotransfection analysis and confocal immunolocalization were used to investigate the significance of one of the identified interactions.
  
  
  RESULTS
  Proliferating cell nuclear antigen (PCNA) was identified as a Prox1 interacting protein. Prox1 interactions with PCNA require the PCNA interacting protein motif (PIP box), located in the Prospero domain of Prox1. Computer modeling of this interaction identified the apparent geometry of this interface which maintains the accessibility of Prox1 to DNA. Prox1 activated the chicken betaB1-crystallin promoter in cotransfection tests as previously reported, while PCNA squelched this transcriptional activation.
  
  
  CONCLUSIONS
  Since PCNA is expressed in the lens epithelium where Prox1 levels are low, while chicken betaB1-crystallin expression activates in lens fibers where Prox1 expression is high and PCNA levels are low, these data suggest that Prox1-PCNA interactions may in part prevent the activation of betaB1-crystallin expression in the lens epithelium.
authors.affiliation: Department of Biological Sciences, University of Delaware, Newark, DE 19716, USA.
authors.forename: Xiaoren
authors.initials: X
authors.lastname: Chen
authors.forename: Tapan P
authors.initials: TP
authors.lastname: Patel
authors.forename: Vladimir I
authors.initials: VI
authors.lastname: Simirskii
authors.forename: Melinda K
authors.initials: MK
authors.lastname: Duncan
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D016466:CHO Cells; D002645:Chickens; D006224:Cricetinae; D003412:Cricetulus; D018398:Homeodomain Proteins / Q000737:chemistry / Q000378:metabolism*; D006801:Humans; D007908:Lens, Crystalline / Q000378:metabolism; D008099:Liver / Q000196:embryology / Q000378:metabolism; D051379:Mice; D008958:Models, Molecular; D008969:Molecular Sequence Data; D018809:Proliferating Cell Nuclear Antigen / Q000737:chemistry / Q000378:metabolism*; D011401:Promoter Regions, Genetic / Q000235:genetics; D011485:Protein Binding; D012097:Repressor Proteins / Q000737:chemistry / Q000378:metabolism*; D014158:Transcription, Genetic; D015533:Transcriptional Activation; D014162:Transfection; D025521:Tumor Suppressor Proteins / Q000737:chemistry / Q000378:metabolism*; D020798:Two-Hybrid System Techniques; D038206:beta-Crystallin B Chain / Q000235:genetics; D000098551:Prospero-Related Homeobox 1 Protein
chemical_list: D018398:Homeodomain Proteins; D018809:Proliferating Cell Nuclear Antigen; D012097:Repressor Proteins; D025521:Tumor Suppressor Proteins; D038206:beta-Crystallin B Chain; D000098551:Prospero-Related Homeobox 1 Protein