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Transcription profiling of mouse embryonic fibroblasts normoxia/hypoxia treatments with mutations in the CH1 domains of p300 and CBP

ABSTRACT: The CH1 (TAZ) domain of the transcriptional coactivators p300 and CBP has been reported to interact with the transcription factor HIF-1alpha and this interaction is thought to be critical for HIF-1alpha target gene expression in response to hypoxia. To determine the requirement for the CH1 domain in hypoxia-responsive gene expression, primary mouse embryonic fibroblasts (MEFs) were generated from e14.5 C57B/6x129/Sv F2 embryos that were either wildtype or bore deletion mutations in the CH1 protein binding domains of both alleles of p300 and one allele of CBP (tri_CH1). Subconfluent MEFs were treated with 21% oxygen (normoxia) or 0.1% oxygen (hypoxia) with 5% carbon dioxide at 37 C in a humid chamber for 6hrs. At the start of treatment, medium was removed and replaced with medium (DMEM+10% FBS+pen-strep+ l-glu) that had been preequilibrated overnight in normoxia or hypoxia as appropriate. Immediately after treatment, cells were lysed in Trizol for RNA extraction. Experiment Overall Design: 4 samples: two genotypes (1 control, 1 mutant); two treatments (normoxia, hypoxia).

ORGANISM(S): Mus musculus

SUBMITTER: Paul Brindle 

PROVIDER: E-GEOD-3195 | biostudies-arrayexpress |

REPOSITORIES: biostudies-arrayexpress

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Two transactivation mechanisms cooperate for the bulk of HIF-1-responsive gene expression.

Kasper Lawryn H LH   Boussouar Fayçal F   Boyd Kelli K   Xu Wu W   Biesen Michelle M   Rehg Jerold J   Baudino Troy A TA   Cleveland John L JL   Brindle Paul K PK  

The EMBO journal 20051020 22

The C-terminal activation domain (C-TAD) of the hypoxia-inducible transcription factors HIF-1alpha and HIF-2alpha binds the CH1 domains of the related transcriptional coactivators CREB-binding protein (CBP) and p300, an oxygen-regulated interaction thought to be highly essential for hypoxia-responsive transcription. The role of the CH1 domain in vivo is unknown, however. We created mutant mice bearing deletions in the CH1 domains (DeltaCH1) of CBP and p300 that abrogate their interactions with t  ...[more]

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