Effect of deletion of 5'HS3 or 5'HS2 of the human beta-globin locus control region on the developmental regulation of globin gene expression in beta-globin locus yeast artificial chromosome transgenic mice.

Peterson, Kenneth R.; Clegg, Christopher H.; Navas, Patrick A.; Norton, Elizabeth J.; Kimbrough, Tyler G.; Stamatoyannopoulos, George

doi:10.1073/pnas.93.13.6605

Cited by 108 publications

(112 citation statements)

References 35 publications

(41 reference statements)

Supporting

Mentioning

Contrasting

Unclassified

Order By: Relevance

“…This result is in agreement with observations that HS2 and HS3 have the strongest enhancer activity (Fraser et al 1993;Peterson et al 1996). In addition, analysis.…”

Section: Validation Of 5c Through Analysis Of the ␤-Globin Locussupporting

confidence: 93%

Chromosome Conformation Capture Carbon Copy (5C): A massively parallel solution for mapping interactions between genomic elements

Dostie¹,

et al. 2006

View full text Add to dashboard Cite

Physical interactions between genetic elements located throughout the genome play important roles in gene regulation and can be identified with the Chromosome Conformation Capture (3C) methodology. 3C converts physical chromatin interactions into specific ligation products, which are quantified individually by PCR. Here we present a high-throughput 3C approach, 3C-Carbon Copy (5C), that employs microarrays or quantitative DNA sequencing using 454-technology as detection methods. We applied 5C to analyze a 400-kb region containing the human ␤-globin locus and a 100-kb conserved gene desert region. We validated 5C by detection of several previously identified looping interactions in the ␤-globin locus. We also identified a new looping interaction in K562 cells between the ␤-globin Locus Control Region and the ␥-␦-globin intergenic region. Interestingly, this region has been implicated in the control of developmental globin gene switching. 5C should be widely applicable for large-scale mapping of cis-and trans-interaction networks of genomic elements and for the study of higher-order chromosome structure.

show abstract

“…This result is in agreement with observations that HS2 and HS3 have the strongest enhancer activity (Fraser et al 1993;Peterson et al 1996). In addition, analysis.…”

Section: Validation Of 5c Through Analysis Of the ␤-Globin Locussupporting

confidence: 93%

Chromosome Conformation Capture Carbon Copy (5C): A massively parallel solution for mapping interactions between genomic elements

Dostie¹,

et al. 2006

View full text Add to dashboard Cite

show abstract

“…The corresponding studies of the endogenous murine LCR have to this point been limited to large deletions encompassing both HS cores and flanking regions. The resulting phenotypes have resembled the milder phenotypes of full HS deletions within transgenic human loci, including moderate suppression of gene expression [14][15][16]26 and little or no effect on formation of the remaining HSs. 17 We have performed a deletion of the core region of 5ЈHS2 of the mouse ␤-globin LCR to more broadly test the hypothesis that deletion of a core is more suppressive than deletion of an entire HS.…”

Section: Discussionmentioning

confidence: 81%

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

Bulger

Bender

et al. 2006

Blood

View full text Add to dashboard Cite

The ␤-globin locus control region (LCR) is a large DNA element that is required for high-level expression of ␤-like globin genes from the endogenous mouse locus or in transgenic mice carrying the human ␤-globin locus. The LCR encompasses 6 DNaseI hypersensitive sites (HSs) that bind transcription factors. These HSs each contain a core of a few hundred base pairs (bp) that has most of the functional activity and exhibits high interspecies sequence homology. Adjoining the cores are 500-to 1000-bp "flanks" with weaker functional activity and lower interspecies homology. Studies of human ␤-globin transgenes and of the endogenous murine locus show that deletion of an entire HS (core plus flanks) moderately suppresses expression. However, human transgenes in which only individual HS core regions were deleted showed drastic loss of expression accompanied by changes in chromatin structure. To address these disparate results, we have deleted the core region of 5HS2 from the endogenous murine ␤-LCR. The phenotype was similar to that of the larger 5HS2 deletion, with no apparent disruption of chromatin structure. These results demonstrate that the greater severity of HS core deletions in comparison to full HS deletions is not a general property of the ␤-LCR. IntroductionLocus control regions (LCRs) are specialized tissue-specific DNA regulatory elements that are functionally defined by their ability to improve position independence and copy number dependence of linked genes in transgenic mice. The ␤-globin LCR was the first identified and has been the most heavily studied. Other LCRs have been identified that are postulated to be important in regulating genes at their endogenous locations. [1][2][3] However, it is unclear whether regulatory activities demonstrated in transgenic studies are relevant for LCRs in their endogenous loci.The human and mouse ␤-globin loci are highly homologous. Both have a series of ␤-like globin genes that are activated in erythroid cells at different developmental stages (reviewed in Stamatoyannopoulos et al 4 ). The human ␤-globin LCR is a group of hypersensitive sites (HSs) located 6 to 22 kb upstream of the embryonic ⑀-globin gene. 5,6 The murine ␤-globin LCR is very similar to the human ␤-globin LCR in its structure, sequence, and activity 7 (Figure 1). The human LCR has been studied predominantly as transgenes in mice or cells, whereas the murine LCR has been predominantly studied by targeted mutation in its endogenous location. Deletion of the murine ␤-globin LCR showed that it is required for high-level expression of all the linked ␤-like globin genes, but developmental timing of gene expression and the chromatin structure of the remaining locus was unaffected by the LCR deletion. 8,9 Each HS contains a very highly conserved core fragment that is 200-to 400-bp long and less conserved but recognizably homologous sequences of roughly 500 to 1000 bp flanking the core ( Figure 1B). The cores from the major HS contain transcription factor binding sites that are generally similar between HSs....

show abstract

“…Although the results of some studies of deletions of individual LCR elements in transgenic human ␤-globin loci have led to the suggestion of stage-specific roles for these elements (23), this conclusion is not true for the murine locus (24)(25)(26) and is unclear for human locus transgenes in general. Deletions of different HSs from human transgenes have parallel effects on developmental regulation, with the greatest decreases in expression occurring at the embryonic and fetal stages (9,(27)(28)(29). These results suggest that any given HS deletion is not associated with a defect in switching but that there is a general difference in sensitivity of the human locus to LCR mutations at different developmental stages.…”

mentioning

confidence: 84%

Promoters of the murine embryonic β-like globin genes Ey and β h1 do not compete for interaction with the β-globin locus control region

Bulger

Roach

et al. 2003

Proc. Natl. Acad. Sci. U.S.A.

View full text Add to dashboard Cite

Mammalian ␤-globin loci contain multiple ␤-like genes that are expressed at different times during development. The murine ␤-globin locus contains two genes expressed during the embryo stage, Ey and ␤h1, and two genes expressed at both the fetal and postnatal stages, ␤-major and ␤-minor. Studies of transgenic human ␤-like globin loci in mice have suggested that expression of one gene at the locus will suppress expression of other genes at the locus. To test this hypothesis we produced mouse lines with deletions of either the Ey or ␤h1 promoter in the endogenous murine ␤-globin locus. Promoter deletion eliminated expression of the mutant gene but did not affect expression of the remaining embryonic gene or the fetal͞adult ␤-globin genes on the mutant allele. These results demonstrate a lack of competitive effects between individual mouse embryonic ␤-globin gene promoters and other genes in the locus. The implication of these findings for models of ␤-globin gene expression are discussed.

show abstract

Effect of deletion of 5'HS3 or 5'HS2 of the human beta-globin locus control region on the developmental regulation of globin gene expression in beta-globin locus yeast artificial chromosome transgenic mice.

Cited by 108 publications

References 35 publications

Chromosome Conformation Capture Carbon Copy (5C): A massively parallel solution for mapping interactions between genomic elements

Chromosome Conformation Capture Carbon Copy (5C): A massively parallel solution for mapping interactions between genomic elements

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

Promoters of the murine embryonic β-like globin genes Ey and β h1 do not compete for interaction with the β-globin locus control region

Contact Info

Product

Resources

About