ABSTRACT
The deoxyribonucleic acid (DNA) damage response (DDR) is a major feature in the maintenance of genome integrity and in the suppression of tumorigenesis. PALB2 (Partner and Localizer of Breast Cancer 2 (BRCA2)) plays an important role in maintaining genome integrity through its role in the Fanconi anemia (FA) and homologous recombination (HR) DNA repair pathways. Since its identification as a BRCA2 interacting partner, PALB2 has emerged as a pivotal tumor suppressor protein associated to hereditary cancer susceptibility to breast and pancreatic cancers. In this review, we discuss how other DDR proteins (such as the kinases Ataxia Telangiectasia Mutated (ATM) and ATM- and Rad3-Related (ATR), mediators BRCA1 (Breast Cancer 1)/BRCA2 and effectors RAD51/DNA Polymerase η (Polη) interact with PALB2 to orchestrate DNA repair. We also examine the involvement of PALB2 mutations in the predisposition to cancer and the role of PALB2 in stimulating error-free DNA repair through the FA/HR pathway.
Subject(s)
DNA Damage , Fanconi Anemia Complementation Group N Protein , Genetic Predisposition to Disease , Genomic Instability , Neoplasms , Recombinational DNA Repair , Animals , Fanconi Anemia Complementation Group N Protein/genetics , Fanconi Anemia Complementation Group N Protein/metabolism , Humans , Neoplasms/genetics , Neoplasms/metabolism , Neoplasms/pathologyABSTRACT
The DNA damage checkpoint (DDC) is an evolutionarily conserved signaling pathway that is crucial to maintain genomic integrity. In response to DNA damage, DDC kinases are rapidly activated and phosphorylate an elaborate network of substrates involved in multiple cellular processes. An important role of the DDC response is to assemble protein complexes. However, for most of the DDC substrates, how the DDC-dependent phosphorylation modulates their network of interactions remains to be established. Here, we present a protocol for the identification of DDC-dependent protein-protein interactions based on Stable Isotope Labeling of Amino acids in Cell culture (SILAC) followed by affinity-tagged protein purification and quantitative mass spectrometry analysis. Based on a model study using Saccharomyces cerevisiae, we provide a method that can be generally applied to study the role of kinases in mediating protein-protein interactions.
Subject(s)
DNA Damage , Saccharomyces cerevisiae Proteins/metabolism , Saccharomyces cerevisiae/metabolism , Tandem Mass Spectrometry/methods , Affinity Labels , Cell Culture Techniques , Chromatography, Liquid/methods , Isotope Labeling , Saccharomyces cerevisiae/geneticsABSTRACT
The platyhelminth parasite Schistosoma mansoni, the causative agent of schistosomiasis, is a dioecious parasite with a complex life cycle that includes two different hosts and two free-living stages. Yet very little is known about the biochemical details connected to these different transitions. In the present work, results will be presented showing the most recent results in S. mansoni regarding the characterization of transcription factors and coactivators that act directly on the transcriptional machinery and those that are involved with chromatin remodeling. It is hoped that the information gathered here may contribute towards the understanding of crucial events in the parasite life cycle. Likewise, the development of new drugs that could interfere with oogenesis and sexual maturation may eventually profit from the information contained herein.
