Deletion of the core region of 5′ HS2 of the mouse β-globin locus control region reveals a distinct effect in comparison with human β-globin transgenes

Hu, Xiao; Bulger, Michael; Bender, Michael A.; Fields, Jennifer; Groudine, Mark; Fiering, Steven

doi:10.1182/blood-2005-06-2308

Cited by 14 publications

(17 citation statements)

References 32 publications

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“…In the ␤-globin locus, six hypersensitive sites (HS1-6) of the LCR, the upstream 5Ј HS-60/-62, and the downstream 3Ј HS1 together form an active chromatin hub (30), where it is widely thought that the LCR and promoters associate with each other. In the ␤-globin LCR, the deletion of HS2, which acts as an enhancer in transient reporter assays, leads to a slight decrease in the expression of the Bmajor and Bminor genes in the fetal liver and adult peripheral blood definitive erythroid cells (31,32). In the present study, we identified six HS sites within the CCR.…”

Section: Discussionmentioning

confidence: 56%

Identification of the Cluster Control Region for the Protocadherin-β Genes Located beyond the Protocadherin-γ Cluster

Yokota

Hirayama

Hirano

et al. 2011

Journal of Biological Chemistry

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The clustered protocadherins (Pcdhs), Pcdh-␣, -␤, and -␥, are transmembrane proteins constituting a subgroup of the cadherin superfamily. Each Pcdh cluster is arranged in tandem on the same chromosome. Each of the three Pcdh clusters shows stochastic and combinatorial expression in individual neurons, thus generating a hugely diverse set of possible cell surface molecules. Therefore, the clustered Pcdhs are candidates for determining neuronal molecular diversity. Here, we showed that the targeted deletion of DNase I hypersensitive (HS) site HS5-1, previously identified as a Pcdh-␣ regulatory element in vitro, affects especially the expression of specific Pcdh-␣ isoforms in vivo. We also identified a Pcdh-␤ cluster control region (CCR) containing six HS sites (HS16, 17, 17, 18, 19, and 20) downstream of the Pcdh-␥ cluster. This CCR comprehensively activates the expression of the Pcdh-␤ gene cluster in cis, and its deletion dramatically decreases their expression levels. Deleting the CCR nonuniformly down-regulates some Pcdh-␥ isoforms and does not affect Pcdh-␣ expression. Thus, the CCR effect extends beyond the 320-kb region containing the Pcdh-␥ cluster to activate the upstream Pcdh-␤ genes. Thus, we concluded that the CCR is a highly specific regulatory unit for Pcdh-␤ expression on the clustered Pcdh genomic locus. These findings suggest that each Pcdh cluster is controlled by distinct regulatory elements that activate their expression and that the stochastic gene regulation of the clustered Pcdhs is controlled by the complex chromatin architecture of the clustered Pcdh locus.

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Section: Discussionmentioning

confidence: 56%

Identification of the Cluster Control Region for the Protocadherin-β Genes Located beyond the Protocadherin-γ Cluster

Yokota

Hirayama

Hirano

et al. 2011

Journal of Biological Chemistry

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“…41 However, deletion of HS2 led to only a 30% decrease in ␤-globin transcription. 41,42 In contrast, KD of PRMT1 has a more drastic effect, more similar to deletion of the entire LCR. The effect is likely because PRMT1 is recruited not only to the LCR but also to the ␤ maj -promoter, and possibly to other genes as well.…”

Section: Discussionmentioning

confidence: 97%

H4R3 methylation facilitates β-globin transcription by regulating histone acetyltransferase binding and H3 acetylation

Patel

et al. 2010

Blood

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Histone modifications play an important role in the process of transcription. However, in contrast to lysine methylation, the role of arginine methylation in chromatin structure and transcription has been underexplored. The globin genes are regulated by a highly organized chromatin structure that juxtaposes the locus control region (LCR) with downstream globin genes. We report here that the targeted recruitment of asymmetric dimethyl H4R3 catalyzed by PRMT1 (protein arginine methyltransferase 1) facilitates histone H3 acetylation on Lys9/Lys14. Dimethyl H4R3 provides a binding surface for P300/ CBP-associated factor (PCAF) and directly enhances histone H3 acetylation in vitro. We show that these active modifications are essential for efficient interactions between the LCR and the ␤ maj -promoter as well as transcription of the ␤-globin gene. Furthermore, knockdown (KD) of PRMT1 by RNA interference in erythroid progenitor cells prevents histone acetylation, enhancer and promoter interaction, and recruitment of transcription complexes to the active ␤-globin promoter. Reintroducing rat PRMT1 into the PRMT1 KD MEL cells rescues PRMT1 binding, ␤-globin transcription, and erythroid differentiation. Taken together, our data suggest that PRMT1-mediated dimethyl H4R3 facilitates histone acetylation and enhancer/ promoter communications, which lead to the efficient recruitment of transcription preinitiation complexes to active promoters. (Blood. 2010;115:2028-2037 IntroductionCovalent modifications of N-terminal histone tails are critically involved in transcriptional activation and repression. 1 The interplay between individual modifications may exert distinct regulatory effects on different gene loci during development and cellular differentiation. For example, H3K9 and H3K27 methylations are generally linked to gene repression, whereas methylation of H3K4 correlates with transcriptionally active euchromatin. 2 However, in the ␤-globin locus H3K9 methylation was also detected in the active globin genes. 3 Arginine methylation of histones is associated with both transcriptional repression and activation. 4 PRMT6-mediated H3R2 dimethylation negatively regulates deposition of H3K4 trimethylation at active promoters, 5 whereas dimethyl H4R3 correlates with transcriptional activation. 6,7 Asymmetric dimethylation of H4R3 residues by protein arginine methyltransferase PRMT1 is essential in vitro and in vivo for the establishment or maintenance of active histone acetylation patterns. 7,8 The interdependence of these modifications appears to be important for the transcription of a p53-dependent reporter gene in a cell-free system with reconstituted chromatin templates. 9 Furthermore, PRMT1 was associated with the activation of HNF4 and HoxA9 genes during tissue development and oncogenesis, respectively. 10, 11 We showed recently that PRMT1 directly interacts with transcription factor USF1 (upstream regulatory factor 1), which has been implicated in chromatin barrier function and ␤-globin gene regulation. [12][13][14][15] The ...

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“…This may explain why small deletions of LCR core regions in transgenic models have a devastating effect on transcription compared with larger deletions, 4,11,12,[38][39][40] but that no such difference is observed at the endogenous locus. 41 …”

Section: Hs Contributions To Lcr Functionmentioning

confidence: 99%

The hypersensitive sites of the murine β-globin locus control region act independently to affect nuclear localization and transcriptional elongation

Bender

Ragoczy

Lee

et al. 2012

Blood

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The ␤-globin locus control region (LCR) is necessary for high-level ␤-globin gene transcription and differentiation-dependent relocation of the ␤-globin locus from the nuclear periphery to the central nucleoplasm and to foci of hyperphosphorylated Pol II "transcription factories" (TFys). To determine the contribution of individual LCR DNaseI hypersensitive sites (HSs) to transcription and nuclear location, in the present study, we compared ␤-globin gene activity and location in erythroid cells derived from mice with deletions of individual HSs, deletions of 2 HSs, and deletion of the whole LCR and found all of the HSs had a similar spectrum of activities, albeit to different degrees. Each HS acts as an independent module to activate expression in an additive manner, and this is correlated with relocation away from the nuclear periphery. In contrast, HSs have redundant activities with respect to association with TFys and the probability that an allele is actively transcribed, as measured by primary RNA transcript FISH. The limiting effect on RNA levels occurs after ␤-globin genes associate with TFys, at which time HSs contribute to the amount of RNA arising from each burst of transcription by stimulating transcriptional elongation. IntroductionAnalysis of naturally occurring and targeted deletions of the endogenous ␤-globin locus control region (LCR) and of transgenic mice has revealed that the LCR is required for high-level ␤-globin transcription in erythroid cells. [1][2][3] The LCR DNase1 hypersensitive sites (HSs) contain similar, albeit distinct sequence motifs, and vary in their activities in vitro and in transgenic mouse assays. [2][3][4][5][6][7][8][9][10] Therefore, each HS may play a unique role in LCR function or may contribute a similar spectrum of activities. Transgenic mice bearing individual LCR HSs, combinations of HSs, or an intact LCR linked to the human ␤-globin locus have led to models in which the LCR HSs interact to form a holocomplex in which the individual HSs 1-4 act synergistically to ensure a permissive chromatin environment and activate expression, whereas 5ЈHSs 5 and 6 act as a chromatin barrier. 5,[11][12][13][14][15][16][17][18][19] In addition, it has been suggested that 5ЈHS2 has a dominant or unique activity compared with other LCR HSs. In contrast to these transgenic studies, our analysis of the transcriptional phenotypes of mice in which each LCR HS was deleted individually from the endogenous locus suggested that each site might contribute additively to LCR-mediated gene activation. [20][21][22] The LCR has also been implicated in the localization of the ␤-globin locus during erythroid differentiation. During differentiation, the ␤-globin locus relocates away from the nuclear periphery and increasingly associates with foci of hyperphosphorylated Pol II "transcription factories" (TFys), where high-level transcription is activated. 23 Deletion of the HSs that comprise the endogenous mouse ␤-globin LCR results in both decreased relocation of the locus to the nucleoplasm and asso...

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Deletion of the core region of 5′ HS2 of the mouse β-globin locus control region reveals a distinct effect in comparison with human β-globin transgenes

Cited by 14 publications

References 32 publications

Identification of the Cluster Control Region for the Protocadherin-β Genes Located beyond the Protocadherin-γ Cluster

Identification of the Cluster Control Region for the Protocadherin-β Genes Located beyond the Protocadherin-γ Cluster

H4R3 methylation facilitates β-globin transcription by regulating histone acetyltransferase binding and H3 acetylation

The hypersensitive sites of the murine β-globin locus control region act independently to affect nuclear localization and transcriptional elongation

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