RhoA belongs to the superfamily of ras-related proteins (38).
Translocation of RhoA and association of translocated RhoA with HSP27 in the particulate fraction, in response to contraction induced by acetylcholine or by ceramide.
inhibit RhoA translocation from the cytosol to the membrane.
Interestingly, RhoA is also known to be instrumental in the kinetics of cyclin D1 expression, specifically in G1 phase of the cell cycle [8].
RhoA G14V (a construct containing a mutation at G14V to render it constitutively active) and RhoA T19N (a construct containing a mutation at T19N, giving it a dominant negative phenotype) constructs were utilized in a cDNA3.1+ vector and were obtained from the Guthrie research institute http://www.cdna.org.
Transient transfection of dominant-negative RhoA (RhoA T19N) and constitutively active (RhoA G14V) constructs were utilised to confirm the regulatory role of Rho in cyclin D1 induction (Fig 3a).
RhoA is a small GTPase that acts as a molecular switch to control a variety of signal transduction pathways in eukaryotes.
In earlier studies from this laboratory that employed differential display and in situ hybridisation, RhoA was indicated as being up-regulated during the stages of early heart development in the developing chick embryo.
To investigate such a mechanism as a possible explanation for the requirement of RhoA in cardiomyocyte differentiation, the P19CL6 inducible cell system over-expressing different forms of RhoA was analysed through real-time PCR to quantify the levels of transcription of genes known to play an important role in early heart development.
The levels of each transcript in transgenic samples (right) were calculated as a ratio of the levels seen in nontransgenic mice, with a mean value of >1 representing induction and a value of <1 representing inhibition of gene expression in transgenic mice relative to nontransgenic mice.
Cardiac and hepatic pathology in RhoA transgenic mice.
the bradycardia in RhoA transgenic mice was not reversed.
Isoprenylcysteine Carboxyl Methyltransferase Modulates Endothelial Monolayer Permeability: Involvement of RhoA Carboxyl ...(Site not responding. Last check: 2007-10-16)
RhoA methylation is presented as the mean±SE of the ratio of methylated RhoA to total RhoA (A).
RhoA in PAECs stably overexpressing GFP or ICMT-GFP by infection
Overexpression of RhoA protein was confirmed by immunoblot analysis (A), and the effects of dominant-negative RhoA on endothelial monolayer permeability were determined by measuring the electrical resistance across the monolayers (B).
RhoA in cardiomyocytes and is involved in sarcomere assembly.
RhoA V14 does not affect (a) GATA-4 protein level or (b) GATA-4 DNA-binding activity, as assessed by Western blot and EMSA, using nuclear extracts of NIH 3T3 cells ectopically expressing either one or both proteins respectively.
RhoA was recently shown to be a potent activator of the p38 mitogen-activated protein kinases (Marinissen et al.
Cultured neonatal cardiomyocytes were transfected with 0.5 µg SK-Luc reporter plasmid together with either the control vector plasmid (1 µg) or expression plasmid encoding RhoA (0.1, 0.25, or 0.5 µg), V14-RhoA (0.1, 0.25, or 0.5 µg), SRFpm1 (0.25 µg), SRFpm1 (0.25 µg) with V14-RhoA (0.5 µg), N19-RhoA (0.5 µg), or C3 transferase (0.25 µg).
The effect of each transfection dose on the activity of the reporter gene was measured in duplicate or triplicate, and the results indicated are representative of three to five independent experiments.
RhoA is required for the clustering of the integrin in fibroblasts through
RhoA- and RhoD-dependent regulatory switch of G{alpha} subunit signaling by PAR-1 receptors in cellular invasion -- NGUYEN et al.
As controls, MDCKT23-DNRac cells were continuously grown in the presence of the DOX repressor (20 ng/ml) to block expression of the dominant-negative form of Rac1 (+DOX).
Lower panel: induction of invasion by TRAP (10 µM) in stably transfected MDCKts.src-DCRhoD cells expressing the constitutively activated form of the RhoA inhibitor RhoD harboring a myc epitope.