Targeted inactivation of Gh/tissue transglutaminase II.
The novel G-protein, G(h)/tissue transglutaminase (TGase II), has both guanosine triphosphatase and Ca(2+)-activated transglutaminase activity and has been implicated in a number of processes including signal transduction, apoptosis, bone ossification, wound healing, and cell adhesion and spreading. To determine the role of G(h) in vivo, the Cre/loxP site-specific recombinase system was used to develop a mouse line in which its expression was ubiquitously inactivated. Despite the absence of G(h) expression and a lack of intracellular TGase activity that was not compensated by other TGases, the Tgm2(-/-) mice were viable, phenotypically normal, and were born with the expected Mendelian frequency. Absence of G(h) coupling to alpha(1)-adrenergic receptor signaling in Tgm2(-/-) mice was demonstrated by the lack of agonist-stimulated [alpha-(32)P]GTP photolabeling of a 74-kDa protein in liver membranes. Annexin-V positivity observed with dexamethasone-induced apoptosis was not different in Tgm2(-/-) thymocytes compared with Tgm2(+/+) thymocytes. However, with this treatment there was a highly significant decrease in the viability (propidium iodide negativity) of Tgm2(-/-) thymocytes. Primary fibroblasts isolated from Tgm2(-/-) mice also showed decreased adherence with culture. These results indicate that G(h) may be importantly involved in stabilizing apoptotic cells before clearance, and in responses such as wound healing that require fibroblast adhesion mediated by extracellular matrix cross-linking.
Molecular Cardiology and Enzyme Research Units, Victor Chang Cardiac Research Institute, Darlinghurst, New South Wales, 2010, Australia.
N
N
Nanda
S E
SE
Iismaa
W A
WA
Owens
A
A
Husain
F
F
Mackay
R M
RM
Graham
D000818:Animals; D001483:Base Sequence; D017931:DNA Primers; D019426:Integrases / Q000235:genetics; D051379:Mice; D018345:Mice, Knockout; D008822:Mice, Transgenic; D010641:Phenotype; D020133:Reverse Transcriptase Polymerase Chain Reaction; D011503:Transglutaminases / Q000037:antagonists & inhibitors* / Q000235:genetics / Q000502:physiology; D014764:Viral Proteins
D017931:DNA Primers; D014764:Viral Proteins; D011503:Transglutaminases; C045073:Cre recombinase; D019426:Integrases