RGD Reference Report - Transcriptional regulation of a contractile gene by mechanical forces applied through integrins in osteoblasts. - Rat Genome Database

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Transcriptional regulation of a contractile gene by mechanical forces applied through integrins in osteoblasts.

Authors: Wang, J  Su, M  Fan, J  Seth, A  McCulloch, CA 
Citation: Wang J, etal., J Biol Chem 2002 Jun 21;277(25):22889-95.
RGD ID: 625414
Pubmed: PMID:11953441   (View Abstract at PubMed)
DOI: DOI:10.1074/jbc.M203130200   (Journal Full-text)

We examined mechanotranscriptional regulation of the contractile gene, alpha-smooth muscle actin (SMA), in osteoblastic cells. Tensile forces were applied through collagen-coated magnetite beads to ROS17/2.8 cells. These cells were desmin-, vimentin+ and expressed low levels of SMA. After force application (480 piconewton/cell), SMA protein and mRNA were increased but beta-actin was unchanged. Beads coated with bovine serum albumin or poly-L-lysine produced no change of SMA. In cells transiently transfected with plasmids containing the SMA promoter fused to beta-galactosidase or green fluorescent protein coding sequences, SMA promoter activity was increased by approximately 60% after 4 h of force, whereas control (Rous sarcoma virus) promoter activity was unaffected. Transfections with beta-galactosidase or green fluorescent protein reporter constructs showed that force-loaded cells exhibited higher beta-galactosidase activity than cells without force. Cytochalasin D and latrunculin B inhibited force-induced increases of SMA promoter activity. Deletion analyses showed that SMA promoter activity was increased approximately 70% after force with a minimal construct containing 155 bp upstream of the translation start site. The force effect on the SMA promoter was abrogated in cells transfected with CArG-B box mutants. Gel mobility shift analyses of nuclear extracts showed strong binding to the CArG-B motif after force. We conclude that the CArG-B box is a force-responsive element in the SMA promoter.



Gene Ontology Annotations    Click to see Annotation Detail View

Biological Process

  
Object SymbolSpeciesTermQualifierEvidenceWithNotesSourceOriginal Reference(s)
Acta2Ratmuscle contraction  IDA  RGD 

Objects Annotated

Genes (Rattus norvegicus)
Acta2  (actin alpha 2, smooth muscle)


Additional Information