AID-Induced Genotoxic Stress Promotes B Cell Differentiation in the Germinal Center via ATM and LKB1 Signaling

Molecular Biology Institute, University of California, Los Angeles, Los Angeles, CA 90095, USA.
Molecular cell (Impact Factor: 14.02). 09/2010; 39(6):873-85. DOI: 10.1016/j.molcel.2010.08.019
Source: PubMed


During an immune response, B cells undergo rapid proliferation and activation-induced cytidine deaminase (AID)-dependent remodeling of immunoglobulin (IG) genes within germinal centers (GCs) to generate memory B and plasma cells. Unfortunately, the genotoxic stress associated with the GC reaction also promotes most B cell malignancies. Here, we report that exogenous and intrinsic AID-induced DNA strand breaks activate ATM, which signals through an LKB1 intermediate to inactivate CRTC2, a transcriptional coactivator of CREB. Using genome-wide location analysis, we determined that CRTC2 inactivation unexpectedly represses a genetic program that controls GC B cell proliferation, self-renewal, and differentiation while opposing lymphomagenesis. Inhibition of this pathway results in increased GC B cell proliferation, reduced antibody secretion, and impaired terminal differentiation. Multiple distinct pathway disruptions were also identified in human GC B cell lymphoma patient samples. Combined, our data show that CRTC2 inactivation, via physiologic DNA damage response signaling, promotes B cell differentiation in response to genotoxic stress.

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Available from: Michael A Damore, Oct 13, 2015
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    • "During CSR, production of DSBs requires the programmed formation of R-loops (Roy et al., 2008) and deoxycytidine deamination mediated by AID (Chaudhuri et al., 2007). The response to DSBs produced by AID activates an ATM-dependent signaling pathway that regulates a network of genes involved in proliferation, B-cell self-renewal, and cell differentiation (Sherman et al., 2010). Interestingly, unscheduled AID-mediated DSBs are implicated in cancer (Park, 2012) even though it is unclear if the link with cancer involves targeting of aberrant R-loop structures. "
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    ABSTRACT: It is well-known that DNA-damaging agents induce genome instability, but only recently have we begun to appreciate that chromosomes are fragile per se and frequently subject to DNA breakage. DNA replication further magnifies such fragility, because it leads to accumulation of single-stranded DNA. Recent findings suggest that chromosome fragility is similarly increased during transcription. Transcripts produced by RNA polymerase II (RNAPII) are subject to multiple processing steps, including maturation of 5' and 3' ends and splicing, followed by transport to the cytoplasm. RNA maturation starts on nascent transcripts and is mediated by a number of diverse proteins and ribonucleoprotein particles some of which are recruited cotranscriptionally through interactions with the carboxy-terminal domain of RNAPII. This coupling is thought to maximize efficiency of pre-mRNA maturation and directly impacts the choice of alternative splice sites. Mounting evidence suggests that lack of coordination among different RNA maturation steps, by perturbing the interaction of nascent transcripts with the DNA template, has deleterious effects on genome stability. Thus, in the absence of proper surveillance mechanisms, transcription could be a major source of DNA damage in cancer. Recent high-throughput screenings in human cells and budding yeast have identified several factors implicated in RNA metabolism that are targets of DNA damage checkpoint kinases: ATM (ataxia telangiectasia mutated) and ATR (ATM-Rad3 related) (Tel1 and Mec1 in budding yeast, respectively). Moreover, inactivation of various RNA processing factors induces accumulation of γH2AX foci, an early sign of DNA damage. Thus, a complex network is emerging that links DNA repair and RNA metabolism. In this review we provide a comprehensive overview of the role played by pre-mRNA processing factors in the cell response to DNA damage and in the maintenance of genome stability.
    Frontiers in Genetics 06/2013; 4:102. DOI:10.3389/fgene.2013.00102
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    • "Rgs1 and Rgs2 were also upregulated in the KI GC B cells. Because of the known role of CREB/CRTC2 target genes in GC B proliferation and of RGS13 in CREB mediated transcription [20], [21], we examined the expression of a number of CREB target genes as well as CREB and CREB co-activators. The KI GC B cells expressed significantly higher levels of CREB1, Crebbp, Crtc2, Ep300, Stk11, Smarca2, and Mta3 (Figure 8B). "
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    ABSTRACT: Germinal centers (GCs) are microanatomic structures that develop in secondary lymphoid organs in response to antigenic stimulation. Within GCs B cells clonally expand and their immunoglobulin genes undergo class switch recombination and somatic hypermutation. Transcriptional profiling has identified a number of genes that are prominently expressed in GC B cells. Among them is Rgs13, which encodes an RGS protein with a dual function. Its canonical function is to accelerate the intrinsic GTPase activity of heterotrimeric G-protein α subunits at the plasma membrane, thereby limiting heterotrimeric G-protein signaling. A unique, non-canonical function of RGS13 occurs following translocation to the nucleus, where it represses CREB transcriptional activity. The functional role of RGS13 in GC B cells is unknown. To create a surrogate marker for Rgs13 expression and a loss of function mutation, we inserted a GFP coding region into the Rgs13 genomic locus. Following immunization GFP expression rapidly increased in activated B cells, persisted in GC B cells, but declined in newly generated memory B and plasma cells. Intravital microscopy of the inguinal lymph node (LN) of immunized mice revealed the rapid appearance of GFP(+) cells at LN interfollicular regions and along the T/B cell borders, and eventually within GCs. Analysis of WT, knock-in, and mixed chimeric mice indicated that RGS13 constrains extra-follicular plasma cell generation, GC size, and GC B cell numbers. Analysis of select cell cycle and GC specific genes disclosed an aberrant gene expression profile in the Rgs13 deficient GC B cells. These results indicate that RGS13, likely acting at cell membranes and in nuclei, helps coordinate key decision points during the expansion and differentiation of naive B cells.
    PLoS ONE 03/2013; 8(3):e60139. DOI:10.1371/journal.pone.0060139 · 3.23 Impact Factor
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    • "There are increasing numbers of reports indicating that the ability of the DNA damage response cascade to modulate cell cycle progression can be used during developmental processes. Some of these processes were linked with limited DNA damage as happens in B cell differentiation (Sherman et al., 2010) or even in the absence of apparent DNA damage, such as in the midblastula transition in Drosophila melanogaster embryos (Sibon et al., 1997) or in the asynchronous division at two-cell-stage Caenorhabditis elegans embryos (Brauchle et al., 2003). The surprising finding that a regulatory cascade involved in DNA damage responses plays a role in a fungal developmental process mirrors these previous results and reinforces the emerging idea that checkpoint cascades may have roles beyond cell surveillance by virtue of their ability to interact with cell cycle machinery. "
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    ABSTRACT: In the phytopathogenic fungus Ustilago maydis, the dikaryotic state dominates the period of growth occurring during the infectious phase. Dikaryons are cells in which two nuclei, one from each parent cell, share a single cytoplasm for a period of time without undergoing nuclear fusion. In fungal cells, maintenance of the dikaryotic state requires an intricate cell division process that often involves the formation of a structure known as the clamp connection as well as the sorting of one of the nuclei to this structure to ensure that each daughter dikaryon inherits a balance of each parental genome. Here, we describe an atypical role of the DNA damage checkpoint kinases Chk1 and Atr1 during pathogenic growth of U. maydis. We found that Chk1 and Atr1 collaborate to control cell cycle arrest during the induction of the virulence program in U. maydis and that Chk1 and Atr1 work together to control the dikaryon formation. These findings uncover a link between a widely conserved signaling cascade and the virulence program in a phytopathogen. We propose a model in which adjustment of the cell cycle by the Atr1-Chk1 axis controls fidelity in dikaryon formation. Therefore, Chk1 and Atr1 emerge as critical cell type regulators in addition to their roles in the DNA damage response.
    The Plant Cell 04/2011; 23(4):1654-65. DOI:10.1105/tpc.110.082552 · 9.34 Impact Factor
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