Expression and localization of plasma transglutaminase factor XIIIA in bone.
Transglutaminases (TGs) are protein crosslinking enzymes involved in cell adhesion and signaling and matrix stabilization and maturation, in many cell types and tissues. We previously described that in addition to transglutaminase 2 (TG2), cultured MC3T3-E1 osteoblasts also express the plasma TG Factor XIIIA (FXIIIA). Here we report on the expression and localization of FXIIIA in bone in vivo and provide confirmatory in vitro data. Immunohistochemistry and in situ hybridization demonstrated that FXIIIA is expressed by osteoblasts and osteocytes in long bones formed by endochondral ossification (femur) and flat bones formed primarily by intramembranous ossification (calvaria and mandible). FXIIIA immunoreactivity was localized to osteoblasts, osteocytes, and the osteoid. RT-PCR analysis revealed FXIIIA expression by both primary osteoblasts and by the MC3T3-E1 osteoblast cell line. Western blot analysis of bone and MC3T3-E1 culture extracts demonstrated that FXIIIA is produced mainly as a small, 37-kDa form. Sequential RT-PCR analysis using overlapping PCR primers spanning the full FXIIIA gene showed that the entire FXIIIA gene is expressed, thus indicating that the 37-kDa FXIIIA is not a splice variant but a product of posttranslational proteolytic processing. Forskolin inhibition of osteoblast differentiation revealed that FXIIIA processing is regulated by the protein kinase A pathway.
Nakano
Yukiko
Y
Faculty of Dentistry, McGill University, Strathcona Bldg. Room M-72, 3640 University Street, Montreal, QC, Canada H3A.
Al-Jallad
Hadil F
HF
Mousa
Aisha
A
Kaartinen
Mari T
MT
D000818:Animals; D001842:Bone and Bones / Q000201:enzymology*; D002454:Cell Differentiation; D002460:Cell Line; D017868:Cyclic AMP-Dependent Protein Kinases / Q000502:physiology; D026122:Factor XIIIa / Q000096:biosynthesis / Q000235:genetics / Q000378:metabolism*; D005269:Femur / Q000201:enzymology; D007150:Immunohistochemistry; D017403:In Situ Hybridization; D051379:Mice; D008807:Mice, Inbred BALB C; D018345:Mice, Knockout; D010006:Osteoblasts / Q000166:cytology / Q000201:enzymology; D010011:Osteocytes / Q000201:enzymology; D012333:RNA, Messenger / Q000096:biosynthesis; D020133:Reverse Transcriptase Polymerase Chain Reaction; D013977:Tibia / Q000201:enzymology; D014020:Tissue Extracts / Q000378:metabolism
D012333:RNA, Messenger; D014020:Tissue Extracts; D026122:Factor XIIIa; D017868:Cyclic AMP-Dependent Protein Kinases