The methylated DNA binding protein-2-H1 (MDBP-2-H1) consists of histone H1 subtypes which are truncated at the C-terminus

Schwarz, Steffen; Heß, Daniel; Jost, Jean-Pierre

doi:10.1093/nar/25.24.5052

Cited by 10 publications

(9 citation statements)

References 28 publications

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“…These sequence-specific DNA-binding proteins show increased binding to these sites in their CpG-methylated form. The sequence specificity of the binding site distinguishes these proteins from methylated DNA-binding proteins, such as MeCP1, MeCP2, DBP-m, and MDBP-2-H1 (22,(32)(33)(34)(35). We demonstrated that the MDBP/RFX could bind to a sequence at the very beginning of the ␣2(I) gene first exon in the rodent and human genomes in a methylation-dependent manner.…”

Section: Discussionmentioning

confidence: 89%

A Methylation-responsive MDBP/RFX Site Is in the First Exon of the Collagen α2(I) Promoter

Sengupta¹,

Ehrlich²,

Smith³

1999

Journal of Biological Chemistry

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DNA methylation inhibits transcription driven by the collagen ␣2(I) promoter and the 5 end of the gene in transient transfection and in vitro transcription assays. DNA-binding proteins in a unique family of ubiquitously expressed proteins, methylated DNA-binding protein (MDBP)/regulatory factor for X box (RFX), form specific complexes with a sequence overlapping the transcription start site of the collagen ␣2(I) gene. Complex formation increased when the CpG site at ؉7 base pairs from the transcription start site was methylated. The identity of the protein was demonstrated by co-migration and cross-competition for a characteristic slowly migrating doublet complex formed on MDBP/RFX recognition sequences and the collagen sequences by band shift assays. A RFX1-specific antibody supershifted the collagen DNA-protein complexes. Furthermore, in vitro translated RFX1 protein formed a specific complex with the collagen sequence that was also supershifted with the RFX1 antibody. MDBP/RFX displayed a higher affinity binding to the collagen sequence if the CpG at ؉7 was mutated in a manner similar to TpG. This same mutation within reporter constructs inhibited transcription in transfection and in vitro transcription assay. These results support the hypothesis that DNA methylationinduced inactivation of collagen ␣2(I) gene transcription is mediated, in part, by increased binding of MDBP/ RFX to the first exon in response to methylation in this region.Type I collagen, the most abundant collagen molecule within the collagen family, normally consists of a heterotrimer of two ␣1(I) chains and one ␣2(I) chain. Synthesis of these genes is often down-regulated upon oncogenic transformation of cells in culture (1-4). We have previously demonstrated down-regulation of the collagen ␣2(I) gene, encoding one of the subunits of Type I collagen in an epithelial-like cell line from rat liver, K16 cells upon their conversion to a tumorigenic line, W8, after treatment with the carcinogen 2-N-(acetoxyacetyl)-aminofluorine (3). The promoter-5Ј region of the ␣2 gene was methylated in the nonexpressing W8 cells and not in the expressing K16 cells (5). Furthermore, reporter gene expression downstream of the 218-bp 1 promoter and the 54-bp 5Ј region of the rat and human collagen ␣2(I) genes was inactivated by in vitro DNA methylation in transient transfection experiments, whereas an analogous expression plasmid with the SV40 early promoter/ enhancer driving expression was not (6). We also demonstrated that a minimal collagen ␣2(I) promoter containing the preinitiation region (Ϫ41 to ϩ54) driving expression of the luciferase reporter gene was inactivated by DNA methylation (7). The inhibition of reporter gene expression was attributable to CpG methylation, specifically of collagen ␣2(I) sequences. However, all the methylation sites were located in the first exon, not in the promoter. DNA methylation in the promoter and 5Ј region of genes often correlates with decreased transcription of vertebrate genes, and many studies indicate that this methyla...

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Section: Discussionmentioning

confidence: 89%

A Methylation-responsive MDBP/RFX Site Is in the First Exon of the Collagen α2(I) Promoter

Sengupta¹,

Ehrlich²,

Smith³

1999

Journal of Biological Chemistry

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“…Such assignment of LHL trypsin‐digested peptides seems to reflect the presence of the H1 and H5 histone sequences in the LHL1 and only H1 histone sequences in the LHL2. Of special interest is a high individual ion sore for the SETAPAPAAEAAPAAAPAPAKA peptide (Table 2) present at the very N‐terminal end of histone H1.c′ (Shannon and Wells, 1987) and MDBP‐2‐H1 (Schwarz et al., 1997). Therefore, it seems that a complex of duck LHL may also contain N‐terminal sequences resembling those of the chicken H1.c′.…”

Section: Resultsmentioning

confidence: 99%

“…In addition to abundant histone H5, the avian erythrocyte linker histones may include up to eight H1 histone variants which can exhibit qualitative and quantitative differences among various species. Linker histone variants H1.z, H1.a, H1.a′, H1.b, H1.b′, H1.c, H1.c′, H1.d and H5 migrate in that order in an acid‐urea polyacrylamide gel (Pałyga, 1991a; Shannon and Wells, 1987). When a gel pattern of linker histones in avian population is analyzed, the allelic isoforms of polymorphic variants with a shifted electrophoretic mobility due to the differences in their molecular weight and/or net charge are occasionally detected.…”

Section: Introductionmentioning

confidence: 99%

Linker histone‐like proteins in Muscovy duck (Cairina moschata L) erythrocyte chromatin

Kowalski

Pałyga

Górnicka-Michalska

2009

Cell Biology International

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Linker Histone-Like proteins (LHL1 and LHL2) were identified within a linker histone complement of Muscovy duck erythrocyte chromatin. Polyacrylamide gel electrophoretic patterns of N-bromosuccinimide-cleaved LHL products as well as liquid chromatography-electrospray-ion trap mass spectrometry analyses of trypsin-digested LHL peptides revealed structural similarity of LHL1 to histone H5 and between LHL2 and histone H1 subtypes. Since the LHL proteins were stable in the presence of 2-mercaptoethanol and dithiothreitol that reduce disulfide bonds, it appeared unlikely that this doublet was a thiol-derived product of linker histones. A loss of LHL1, with a concomitant maintenance of LHL2 after treatment with dilute alkali, seems to suggest that they might represent disparate protein conjugates resulting from linker histone modifications through ester linkages.

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“…Histone H1 colocalized with 5‐methylcytosine (Ball et al ., 1983) and bound preferentially to CpG‐methylated DNA irrespective of DNA sequence (Jost and Hofsteenge, 1992; McArthur and Thomas, 1996). Several reports have established a positive correlation between DNA methylation, binding of histone H1 and transcriptional repression (Levine et al ., 1993; Johnson et al ., 1995; Schwarz et al ., 1997). In contrast with these observations, it was reported that Histone H1 has no preference for methylated DNA (Campoy et al ., 1995) and no role in methylation‐associated gene silencing (Barra et al ., 2000).…”

Section: Discussionmentioning

confidence: 99%

Sensing DNA methylation in the protozoan parasite Entamoeba histolytica

Lavi¹,

Isakov²,

Harony³

et al. 2006

Molecular Microbiology

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SummaryIn the protozoan parasite Entamoeba histolytica, 5-methylcytosine (m5C) was found predominantly in repetitive elements. Its formation is catalysed by Ehmeth, a DNA methyltransferase that belongs to the Dnmt2 subfamily. Here we describe a 32 kDa nuclear protein that binds in vitro with higher affinity to the methylated form of a DNA encoding a reverse transcriptase of an autonomous non-long-terminal repeat retrotransposon (RT LINE) compared with the non-methylated RT LINE. This protein, named E. histolytica-methylated LINE binding protein (EhMLBP), was purified from E. histolytica nuclear lysate, identified by mass spectrometry, and its corresponding gene was cloned. EhMLBP corresponds to a gene of unknown function that shares strong homology with putative proteins present in Entamoeba dispar and Entamoeba invadens. In contrast, the homology dropped dramatically when nonEntamoebidae sequences were considered and only a weak sequence identity was found with Trypanosoma and several prokaryotic histone H1. Recombinant EhMLBP showed the same binding preference for methylated RT LINE as the endogenous EhMLBP. Deletion mapping analysis localized the DNA binding region at the C-terminal part of the protein. This region is sufficient to assure the binding to methylated RT LINE with high affinity. Western blot and immunofluorescence microscopy, using an antibody raised against EhMLBP, showed that it has a nuclear localization. Chromatin immunoprecipitation (ChIP) confirmed that EhMLBP interacts with RT LINE in vivo. Finally, we showed that EhMLBP can also bind rDNA episome, a DNA that is methylated in the parasite. This suggests that EhMLBP may serve as a sensor of methylated repetitive DNA. This is the first report of a DNA-methylated binding activity in protozoa.

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The methylated DNA binding protein-2-H1 (MDBP-2-H1) consists of histone H1 subtypes which are truncated at the C-terminus

Cited by 10 publications

References 28 publications

A Methylation-responsive MDBP/RFX Site Is in the First Exon of the Collagen α2(I) Promoter

A Methylation-responsive MDBP/RFX Site Is in the First Exon of the Collagen α2(I) Promoter

Linker histone‐like proteins in Muscovy duck (Cairina moschata L) erythrocyte chromatin

Sensing DNA methylation in the protozoan parasite Entamoeba histolytica

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