Florence Cammas

Transcription cofactors TRIM24, TRIM28, and TRIM33 associate to form regulatory complexes that suppress murine hepatocellular carcinoma

TRIM24 (TIF1α), TRIM28 (TIF1β), and TRIM33 (TIF1γ) are three related cofactors belonging to the tripartite motif superfamily that interact with distinct transcription factors. TRIM24 interacts with the liganded retinoic acid (RA) receptor to repress its transcriptional activity. Germ line inactivation of TRIM24 in mice deregulates RA-signaling in hepatocytes leading to the development of hepatocellular carcinoma (HCC). Here we show that TRIM24 can be purified as at least two macromolecular complexes comprising either TRIM33 or TRIM33 and TRIM28. Somatic hepatocyte-specific inactivation of TRIM24, TRIM28, or TRIM33 all promote HCC in a cell-autonomous manner in mice. Moreover, HCC formation upon TRIM24 inactivation is strongly potentiated by further loss of TRIM33. These results demonstrate that the TIF1-related subfamily of TRIM proteins interact both physically and functionally to modulate HCC formation in mice.

KAP1-Mediated Epigenetic Repression in the Forebrain Modulates Behavioral Vulnerability to Stress

KAP1 is an essential cofactor of KRAB-zinc finger proteins, a family of vertebrate-specific epigenetic repressors of largely unknown functions encoded in the hundreds by the mouse and human genomes. Here, we report that KAP1 is expressed at high levels and necessary for KRAB-mediated repression in mature neurons of the mouse brain. Mice deleted for KAP1 in the adult forebrain exhibit heightened levels of anxiety-like and exploratory activity and stress-induced alterations in spatial learning and memory. In the hippocampus, a small number of genes are dysregulated, including some imprinted genes. Chromatin analyses of the promoters of two genes markedly upregulated in knockout mice reveal decreased histone 3 K9-trimethylation and increased histone 3 and histone 4 acetylation. We propose a model in which the tethering of KAP1-associated chromatin remodeling factors via KRAB-ZFPs epigenetically controls gene expression in the hippocampus, thereby conditioning responses to behavioral stress.

Epigenetic tethering of AID to the donor switch region during immunoglobulin class switch recombination.

A complex of KAP1 and HP1 is needed to tether AID to the H3K9me3-marked donor switch region during CSR.

Primer Binding Site-Dependent Restriction of Murine Leukemia Virus Requires HP1 Binding by TRIM28

ABSTRACT TRIM28 is a transcriptional corepressor which is required for primer binding site (PBS)-dependent restriction of murine leukemia virus (MLV) replication in embryonic stem and embryonic carcinoma (EC) cells. PBS-dependent restriction of MLV leads to transcriptional silencing of the integrated provirus and has been shown to correlate with TRIM28-mediated recruitment of HP1 to the silenced loci. Here we show, using a cell line with a point mutation in the HP1 binding domain of TRIM28, that interaction with HP1 is absolutely required for the PBS-dependent restriction of MLV in the F9 EC cell line.

The histone subcode: poly(ADP-ribose) polymerase-1 (Parp-1) and Parp-2 control cell differentiation by regulating the transcriptional intermediary factor TIF1beta and the heterochromatin protein HP1alpha.

Recent advances reveal emerging unique functions of poly(ADP‐ribose) polymerase‐1 (Parp‐1) and Parp‐2 in heterochromatin integrity and cell differentiation. However, the chromatin‐mediated molecular and cellular events involved remain elusive. Here we describe specific physical and functional interactions of Parp‐1 and Parp‐2 with the transcriptional intermediary factor (TlF1β) and the heterochromatin proteins (HP1) that affect endodermal differentiation. We show that Parp‐2 binds to TlF1β with high affinity both directly and through HP1α. Both partners colocalize at pericentric heterochromatin in primitive endoderm‐like cells. Parp‐2 also binds to HP1β but not to HP1γ. In contrast Parp‐1 binds weakly to TlF1β and HP1β only. Both Parps selectively poly(ADP‐ribosyl)ate HP1α. Using shRNA approaches, we provide evidence for distinct participation of both Parps in endodermal differentiation. Whereas Parp‐2 and its activity are required for the relocation of TlF1β to heterochromatic foci during primitive endodermal differentiation, Parp‐1 and its activity modulate TlF1β‐HP1α association with consequences on parietal endodermal differentiation. Both Parps control TlF1β transcriptional activity. In addition, this work identifies both Parps as new modulators of the HP1‐mediated subcode histone.— Quénet, D., Gasser, V., Fouillen, L., Cammas, F., Sanglier‐Cianferani, S., Losson, R., Dantzer, F. The histone subcode: poly(ADP‐ribose) polymerase‐1 (Parp‐1) and Parp‐2 control cell differentiation by regulating the transcriptional intermediary factor TIF1β and the heterochromatin protein HPlα. FASEB J. 22, 3853–3865 (2008)

TRIM28 Represses Transcription of Endogenous Retroviruses in Neural Progenitor Cells

SUMMARY TRIM28 is a corepressor that mediates transcriptional silencing by establishing local heterochromatin. Here, we show that deletion of TRIM28 in neural progenitor cells (NPCs) results in high-level expression of two groups of endogenous retroviruses (ERVs): IAP1 and MMERVK10C. We find that NPCs use TRIM28-mediated histone modifications to dynamically regulate transcription and silencing of ERVs, which is in contrast to other somatic cell types using DNA methylation. We also show that derepression of ERVs influences transcriptional dynamics in NPCs through the activation of nearby genes and the expression of long noncoding RNAs. These findings demonstrate a unique dynamic transcriptional regulation of ERVs in NPCs. Our results warrant future studies on the role of ERVs in the healthy and diseased brain.

TIF1β association with HP1 is essential for post-gastrulation development, but not for Sertoli cell functions during spermatogenesis

TIF1β is an essential mammalian transcriptional corepressor. It interacts with the heterochromatin proteins HP1 through a highly conserved motif, the HP1box, and we have previously shown that this interaction is essential for the differentiation of F9 cells to occur. Here we address the in vivo functions of the TIF1β-HP1 interaction, by generating mice in which the TIF1β HP1box is mutated, leading to the loss of TIF1β interaction with HP1. The effects of the mutation were monitored in two instances, where TIF1β is known to play key roles: early embryonic development and spermatogenesis. We find that mutating the HP1box of TIF1β disrupts embryonic development soon after gastrulation. This effect is likely caused by the misexpression of TIF1β targets that regulate mitotic progression and pluripotency. In contrast, in Sertoli cells, we found that the absence of TIF1β but not its mutation in the HP1box leads to a clear defect of spermatogenesis characterized by a failure of spermatid release and a testicular degeneration. These data show that the interaction between TIF1β and HP1 is essential for some but not all TIF1β functions in vivo. Furthermore, we observed that TIF1β is dispersed through the nucleoplasm of E7.0 embryos, whereas it is mainly associated with pericentromeric heterochromatin of E8.5 embryos and of Sertoli cells, an association that is lost upon TIF1β HP1box mutation. Altogether, these data provide strong evidence that nuclear organization plays key roles during early embryonic development.

In mammalian foetal testes, SOX9 regulates expression of its target genes by binding to genomic regions with conserved signatures

Abstract In mammalian embryonic gonads, SOX9 is required for the determination of Sertoli cells that orchestrate testis morphogenesis. To identify genetic networks directly regulated by SOX9, we combined analysis of SOX9-bound chromatin regions from murine and bovine foetal testes with sequencing of RNA samples from mouse testes lacking Sox9. We found that SOX9 controls a conserved genetic programme that involves most of the sex-determining genes. In foetal testes, SOX9 modulates both transcription and directly or indirectly sex-specific differential splicing of its target genes through binding to genomic regions with sequence motifs that are conserved among mammals and that we called ‘Sertoli Cell Signature’ (SCS). The SCS is characterized by a precise organization of binding motifs for the Sertoli cell reprogramming factors SOX9, GATA4 and DMRT1. As SOX9 biological role in mammalian gonads is to determine Sertoli cells, we correlated this genomic signature with the presence of SOX9 on chromatin in foetal testes, therefore equating this signature to a genomic bar code of the fate of foetal Sertoli cells. Starting from the hypothesis that nuclear factors that bind to genomic regions with SCS could functionally interact with SOX9, we identified TRIM28 as a new SOX9 partner in foetal testes.

TRIM Involvement in Transcriptional Regulation

Members of the tripartite motif (TRIM) protein family are found in all multicellular eukaryotes and function in a wide range of cellular processes such as cell cycle regulation, differentiation, development, oncogenesis and viral response. Over the past few years, several TRIM proteins have been reported to control gene expression through regulation of the transcriptional activity of numerous sequence-specific transcription factors. These proteins include the transcriptional intermediary factor 1 (TIF1) regulators, the promyelocytic leukemia tumor suppressor PML and the RET finger protein (RFP). In this chapter, we will consider the molecular interactions made by these TRIM proteins and will attempt to clarify some of the molecular mechanisms underlying their regulatory effect on transcription.

Correlation of the exon/intron organization to the conserved domains of the mouse transcriptional corepressor TIF1β ☆

TIF1beta, a member of the transcriptional intermediary factor 1 family, has been reported to function as a corepressor for the large class of KRAB domain-containing zinc finger proteins of the Krüppel type. In this study, we report the genomic organization and nucleotide sequence of the mouse TIF1beta gene. This gene comprises 17 coding exons located within 7 kb of genomic DNA. Exon sizes vary from 37 bp (exon 10) to 901 bp (exon 1), and intron sizes range from 71 bp to 1843 bp. All introns have the conserved GT and AG dinucleotides present at the donor and acceptor sites, respectively. The functional/homology regions of the TIF1beta protein are encoded by distinct exons. The amino-terminal RING finger is encoded by two exons interrupted by a small intron. The B boxes lie within individual exons. Similarly to the RING finger, the PHD finger is encoded by two exons. Three exons constitute the carboxy-terminal bromodomain, and their position correlates well with the secondary structure elements of the domain as predicted by computer modeling. Taken together, these results will facilitate the genetic manipulation of TIF1beta for future in vivo structure-function studies.