title: A role for the RNA pol II-associated PAF complex in AID-induced immune diversification.
abstract: Antibody diversification requires the DNA deaminase AID to induce DNA instability at immunoglobulin (Ig) loci upon B cell stimulation. For efficient cytosine deamination, AID requires single-stranded DNA and needs to gain access to Ig loci, with RNA pol II transcription possibly providing both aspects. To understand these mechanisms, we isolated and characterized endogenous AID-containing protein complexes from the chromatin of diversifying B cells. The majority of proteins associated with AID belonged to RNA polymerase II elongation and chromatin modification complexes. Besides the two core polymerase subunits, members of the PAF complex, SUPT5H, SUPT6H, and FACT complex associated with AID. We show that AID associates with RNA polymerase-associated factor 1 (PAF1) through its N-terminal domain, that depletion of PAF complex members inhibits AID-induced immune diversification, and that the PAF complex can serve as a binding platform for AID on chromatin. A model is emerging of how RNA polymerase II elongation and pausing induce and resolve AID lesions.
authors.affiliation: DNA Editing Laboratory, London Research Institute, South Mimms EN6 3LD, England, UK.
authors.forename: Katharina L
authors.initials: KL
authors.lastname: Willmann
authors.forename: Sara
authors.initials: S
authors.lastname: Milosevic
authors.forename: Siim
authors.initials: S
authors.lastname: Pauklin
authors.forename: Kerstin-Maike
authors.initials: KM
authors.lastname: Schmitz
authors.forename: Gopinath
authors.initials: G
authors.lastname: Rangam
authors.forename: Maria T
authors.initials: MT
authors.lastname: Simon
authors.forename: Sarah
authors.initials: S
authors.lastname: Maslen
authors.forename: Mark
authors.initials: M
authors.lastname: Skehel
authors.forename: Isabelle
authors.initials: I
authors.lastname: Robert
authors.forename: Vincent
authors.initials: V
authors.lastname: Heyer
authors.forename: Ebe
authors.initials: E
authors.lastname: Schiavo
authors.forename: Bernardo
authors.initials: B
authors.lastname: Reina-San-Martin
authors.forename: Svend K
authors.initials: SK
authors.lastname: Petersen-Mahrt
mesh_terms: D000818:Animals; D000916:Antibody Diversity; D001402:B-Lymphocytes / Q000378:metabolism*; D015153:Blotting, Western; D045744:Cell Line, Tumor; D002843:Chromatin / Q000235:genetics / Q000378:metabolism; D003564:Cytidine Deaminase / Q000235:genetics / Q000378:metabolism*; D004268:DNA-Binding Proteins / Q000235:genetics / Q000378:metabolism; D057809:HEK293 Cells; D006367:HeLa Cells; D006609:High Mobility Group Proteins / Q000235:genetics / Q000378:metabolism; D006801:Humans; D017578:Immunoglobulin Class Switching; D007136:Immunoglobulins / Q000235:genetics; D047468:Immunoprecipitation; D009687:Nuclear Proteins / Q000235:genetics / Q000378:metabolism*; D011485:Protein Binding; D034622:RNA Interference; D014157:Transcription Factors / Q000235:genetics / Q000378:metabolism; D035602:Transcriptional Elongation Factors / Q000235:genetics / Q000378:metabolism; D000098830:AICDA (Activation-Induced Cytidine Deaminase)
chemical_list: D002843:Chromatin; D004268:DNA-Binding Proteins; D006609:High Mobility Group Proteins; D007136:Immunoglobulins; D009687:Nuclear Proteins; C501972:PAF1 protein, human; C073537:SSRP1 protein, human; C112224:SUPT5H protein, human; C100927:SUPT6H protein, human; D014157:Transcription Factors; D035602:Transcriptional Elongation Factors; D000098830:AICDA (Activation-Induced Cytidine Deaminase); D003564:Cytidine Deaminase