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Regulated transcription of human matrix metalloproteinase 13 (MMP13) and interleukin -1B (IL 1B) genes in chondrocytes depends on methylation of specific proximal promoter CpG sites

Regulated transcription of human matrix metalloproteinase 13 (MMP13) and interleukin -1B (IL 1B) genes in chondrocytes depends on methylation of specific proximal promoter CpG sites
Regulated transcription of human matrix metalloproteinase 13 (MMP13) and interleukin -1B (IL 1B) genes in chondrocytes depends on methylation of specific proximal promoter CpG sites
The role of DNA methylation in the regulation of catabolic genes such as MMP13 and IL1B, which have sparse CpG islands, is poorly understood in the context of musculoskeletal diseases. We report that demethylation of specific CpG sites at ?110 bp and ?299 bp of the proximal MMP13 and IL1B promoters, respectively, detected by in situ methylation analysis of chondrocytes obtained directly from human cartilage, strongly correlated with higher levels of gene expression. The methylation status of these sites had a significant impact on promoter activities in chondrocytes, as revealed in transfection experiments with site-directed CpG mutants in a CpG-free luciferase reporter. Methylation of the ?110 and ?299 CpG sites, which reside within a hypoxia-inducible factor (HIF) consensus motif in the respective MMP13 and IL1B promoters, produced the most marked suppression of their transcriptional activities. Methylation of the ?110 bp CpG site in the MMP13 promoter inhibited its HIF-2?-driven transactivation and decreased HIF-2? binding to the MMP13 proximal promoter in chromatin immunoprecipitation assays. In contrast to HIF-2?, MMP13 transcriptional regulation by other positive (RUNX2, AP-1, ELF3) and negative (Sp1, GATA1, and USF1) factors was not affected by methylation status. However, unlike the MMP13 promoter, IL1B was not susceptible to HIF-2? transactivation, indicating that the ?299 CpG site in the IL1B promoter must interact with other transcription factors to modulate IL1B transcriptional activity. Taken together, our data reveal that the methylation of different CpG sites in the proximal promoters of the human MMP13 and IL1B genes modulates their transcription by distinct mechanisms.
cartilage, DNA methylation, hypoxia-inducible factor (HIF), interleukin, matrix metalloproteinase (MMP), transcription promoter
0021-9258
10061-10072
Hashimoto, Ko
19b8f3be-d539-4579-a6cb-936ed0243b27
Otero, Miguel
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Imagawa, Kei
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de Andres, Maria C.
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Coico, Jonathan M.
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Roach, Helmtrud I.
ca2ff1f4-1ada-4c56-9097-cd27ca4d199e
Oreffo, Richard O.C.
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Marcu, Kenneth B.
b7ff235f-4447-4e55-8f1d-43bbf3c20375
Goldring, Mary B.
67cbd000-e36a-4b82-bc50-f3c5a8d292da
Hashimoto, Ko
19b8f3be-d539-4579-a6cb-936ed0243b27
Otero, Miguel
3f7ee13f-d9cf-495e-b184-e9896d32c8cf
Imagawa, Kei
cfdeef65-8259-4f0c-943a-0d439aab3193
de Andres, Maria C.
51b4d6c7-cad3-4818-b27a-2aa86c8656e0
Coico, Jonathan M.
f4c035ff-2b86-454f-8d32-7cd116087181
Roach, Helmtrud I.
ca2ff1f4-1ada-4c56-9097-cd27ca4d199e
Oreffo, Richard O.C.
ff9fff72-6855-4d0f-bfb2-311d0e8f3778
Marcu, Kenneth B.
b7ff235f-4447-4e55-8f1d-43bbf3c20375
Goldring, Mary B.
67cbd000-e36a-4b82-bc50-f3c5a8d292da

Hashimoto, Ko, Otero, Miguel, Imagawa, Kei, de Andres, Maria C., Coico, Jonathan M., Roach, Helmtrud I., Oreffo, Richard O.C., Marcu, Kenneth B. and Goldring, Mary B. (2013) Regulated transcription of human matrix metalloproteinase 13 (MMP13) and interleukin -1B (IL 1B) genes in chondrocytes depends on methylation of specific proximal promoter CpG sites. The Journal of Biological Chemistry, 288 (14), 10061-10072. (doi:10.1074/jbc.M112.421156). (PMID:23417678)

Record type: Article

Abstract

The role of DNA methylation in the regulation of catabolic genes such as MMP13 and IL1B, which have sparse CpG islands, is poorly understood in the context of musculoskeletal diseases. We report that demethylation of specific CpG sites at ?110 bp and ?299 bp of the proximal MMP13 and IL1B promoters, respectively, detected by in situ methylation analysis of chondrocytes obtained directly from human cartilage, strongly correlated with higher levels of gene expression. The methylation status of these sites had a significant impact on promoter activities in chondrocytes, as revealed in transfection experiments with site-directed CpG mutants in a CpG-free luciferase reporter. Methylation of the ?110 and ?299 CpG sites, which reside within a hypoxia-inducible factor (HIF) consensus motif in the respective MMP13 and IL1B promoters, produced the most marked suppression of their transcriptional activities. Methylation of the ?110 bp CpG site in the MMP13 promoter inhibited its HIF-2?-driven transactivation and decreased HIF-2? binding to the MMP13 proximal promoter in chromatin immunoprecipitation assays. In contrast to HIF-2?, MMP13 transcriptional regulation by other positive (RUNX2, AP-1, ELF3) and negative (Sp1, GATA1, and USF1) factors was not affected by methylation status. However, unlike the MMP13 promoter, IL1B was not susceptible to HIF-2? transactivation, indicating that the ?299 CpG site in the IL1B promoter must interact with other transcription factors to modulate IL1B transcriptional activity. Taken together, our data reveal that the methylation of different CpG sites in the proximal promoters of the human MMP13 and IL1B genes modulates their transcription by distinct mechanisms.

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Published date: 5 April 2013
Keywords: cartilage, DNA methylation, hypoxia-inducible factor (HIF), interleukin, matrix metalloproteinase (MMP), transcription promoter
Organisations: Human Development & Health

Identifiers

Local EPrints ID: 355820
URI: http://eprints.soton.ac.uk/id/eprint/355820
ISSN: 0021-9258
PURE UUID: 42848960-42f6-4e39-8570-cdfda2dad72f
ORCID for Richard O.C. Oreffo: ORCID iD orcid.org/0000-0001-5995-6726

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Date deposited: 05 Sep 2013 14:09
Last modified: 28 Oct 2023 01:46

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Contributors

Author: Ko Hashimoto
Author: Miguel Otero
Author: Kei Imagawa
Author: Maria C. de Andres
Author: Jonathan M. Coico
Author: Helmtrud I. Roach
Author: Kenneth B. Marcu
Author: Mary B. Goldring

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