Promoter-2-derived expression in bovine granulosa cells coincides with gene-specific DNA hypo-methylation

Vanselow, Jens; Pöhland, Ralf; Fürbaß, Rainer

doi:10.1016/j.mce.2005.01.007

Cited by 40 publications

(32 citation statements)

References 28 publications

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“…PCR amplification will eventually replace uracil with thymine (T). The bisulphite sequencing protocol was essentially as described (Vanselow et al 2005). The modified target sequences were amplified by PCR with primers that specifically bind to the plus strand of the corresponding genomic regions.…”

Section: Electrophoretic Mobility Shift Assay (Emsa)mentioning

confidence: 99%

“…Primers used to amplify 453 bp from promoter 2 of the bovine Cyp19 gene (used as a control, cf. Vanselow et al 2005) were 5 0 -TGGA-GAAGTTTATTTTTTGGGGGAAGT (forward) and 5 0 -TCCCCAAAATATACA-TTTAAAACTCCT (reverse). PCR was performed in 25 ml reaction volume with 0 .…”

Section: Electrophoretic Mobility Shift Assay (Emsa)mentioning

confidence: 99%

“…The sequence gel files were further processed and evaluated with the PHRED software, version 0.020425 (53;54) in order to quantify the C/T polymorphic sites (Vanselow et al 2005). The percentage of methylation was calculated as the portion of the C peak area within each polymorphic CpG site [% methylationZ(peak area of C/peak area of CCpeak area of T)!100)].…”

Section: Electrophoretic Mobility Shift Assay (Emsa)mentioning

confidence: 99%

“…Thus, the infectionrelated increased degree of methylation resulted in chromatin condensation around the CoRE. To examine, if the infection-related alterations in the percentage of methylated DNA molecules are specific for the aS1-casein, we measured in these udder samples the degree of methylation at three CpG sites on the promoter 2 of the Cyp19 aromatase-encoding gene (Vanselow et al 2005). The Cyp19 aromatase gene is not known to be expressed in udder or to be lactationally regulated.…”

Section: Dna-methylation In the Vicinity Of The Stat Factorbinding Simentioning

confidence: 99%

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DNA-remethylation around a STAT5-binding enhancer in the αS1-casein promoter is associated with abrupt shutdown of αS1-casein synthesis during acute mastitis

Vanselow

Yang²,

Herrmann³

et al. 2006

Journal of Molecular Endocrinology

109

114

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Prolactin stimulates the expression of milk genes during lactation through the activation of STAT5 transcription factors, which subsequently bind to their cognate target sequence on the promoters. Demethylation of 5methylCpG dinucleotides permits the tissue-specific accessibility of transcription factor-binding sites during development, but remethylation has not been shown to contribute to acute suppression of gene expression. We characterize functionally a novel STAT5-binding lactational enhancer in the far upstream promoter (wK10 kbp) of the bovine aS1-casein-encoding gene. This promoter area is hypomethylated in the lactating udder only. Remethylation of this area accompanies an experimentally elicited acute shutdown of casein synthesis in fully lactating cows, whose udder quarters have experimentally been infected with a pathogenic E. coli strain. Within 24 h after infection, the relevant promoter area was remethylated from 10% of the DNA molecules in the uninfected control quarters to w50% in the infected quarters, the typical values for fully lactating and not lactating udders respectively. Increased methylation resulted in tighter chromatin packing. Concomitantly, the aS1-casein mRNA concentration dropped to w50% while the protein synthesis was shut down to w2 . 5% in the infected quarters, alone. The methylation status of the promoter from a not lactationally regulated gene was unaltered, and the distal aS1-casein promoter was not remethylated in udder quarters with subclinical Staphylococcus aureus infections featuring sustained casein synthesis. Hence, infection-related remethylation of the aS1-casein promoter and chromatin remodelling serves as an acute, spatially restricted regulatory mechanism, which might insulate the promoter against the systemically unchanged high levels of circulating prolactin. This provides a rare example for an acute regulatory significance of CpG methylation.

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Section: Electrophoretic Mobility Shift Assay (Emsa)mentioning

confidence: 99%

Section: Electrophoretic Mobility Shift Assay (Emsa)mentioning

confidence: 99%

Section: Electrophoretic Mobility Shift Assay (Emsa)mentioning

confidence: 99%

Section: Dna-methylation In the Vicinity Of The Stat Factorbinding Simentioning

confidence: 99%

See 2 more Smart Citations

DNA-remethylation around a STAT5-binding enhancer in the αS1-casein promoter is associated with abrupt shutdown of αS1-casein synthesis during acute mastitis

Vanselow

Yang²,

Herrmann³

et al. 2006

Journal of Molecular Endocrinology

109

114

View full text Add to dashboard Cite

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“…3 Expression of the gene is controlled by paracrine and endocrine factors and also by DNA methylation. 4 Aromatase activity is key during follicular development as it controls part of follicular differentiation and the exchange between the ovary and the brain. The estradiol concentration is linked to expression of CYP19A1, which encodes the limiting enzyme of the steroidogenic cascade.…”

Section: Introductionmentioning

confidence: 99%

A Genetical Genomics Methodology to Identify Genetic Markers of a Bovine Fertility Phenotype Based on CYP19A1 Gene Expression

Guillemin

Dufort²,

Sirard³

2015

Gene Expression to Genetical Genomics

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ABSTRACT:With the decrease in fertility in dairy cow, the interest towards means to control this variable through genetic selection is growing. One of the most important factors controlling follicular maturity and timely ovulation is the aromatase enzyme, which is encoded by the CYP19A1 gene. The activity of this enzyme is potentially the limiting factor in postpartum fertility. In this study, we developed a methodology, based on genetical genomics, to model the aromatase expression profile from granulosa cell samples. The transcriptomic expression profiles obtained were used to identify 355 genes or isoforms potentially associated with the regulation of aromatase. From those genes, 23,388 single-nucleotide polymorphism (SNPs) in the genome of Holstein cows were identified. Results showed that some SNPs (on KRT8, LHCGR, CREB, ANXA1, and on CYP19A1 itself) were relevant to aromatase expression and the model generated could predict 44% of the observed phenotype. This study demonstrated the value of genetical genomics to generate better biomarkers for the dairy industry.

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Bovine Epigenetics and Epigenomics

Tian

2012

Bovine Genomics

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Promoter-2-derived expression in bovine granulosa cells coincides with gene-specific DNA hypo-methylation

Cited by 40 publications

References 28 publications

DNA-remethylation around a STAT5-binding enhancer in the αS1-casein promoter is associated with abrupt shutdown of αS1-casein synthesis during acute mastitis

DNA-remethylation around a STAT5-binding enhancer in the αS1-casein promoter is associated with abrupt shutdown of αS1-casein synthesis during acute mastitis

A Genetical Genomics Methodology to Identify Genetic Markers of a Bovine Fertility Phenotype Based on CYP19A1 Gene Expression

Bovine Epigenetics and Epigenomics

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