The Sp1 and CBF/NF-Y Transcription Factors Cooperatively Regulate the Mouse Pro-alpha3(V) Collagen Gene (Col5a3) in Osteoblastic Cells

The purpose of this study was to clarify the mechanism responsible for the transcriptional regulation of the mouse Col5a3 gene in osteoblastic cells. Transient transfection into rat osteosarcoma ROS17/2.8 cells demonstrated that a region from nucleotides 337 to 1 was involved in the transcriptional activity of the Col5a3 gene. An electrophoretic mobility shift assay showed that Sp1/Sp3 and CBF/NF-Y bound to a GC-rich domain (194/186) and a CCAAT box (134/130) in the Col5a3 gene, respectively. Introduction of mutations or deletion into a GC-rich domain, the CCAAT box, or both elements decreased the transcription activity. Overexpression of Sp1 increases the transcription activity and interferes with Sp family binding to the GC-rich domain to decrease promoter activity. Therefore, the transcription of the mouse Col5a3 gene is cooperatively regulated by Sp1 and CBF/NF-Y in osteoblastic cells.</p

Multifactor complex containing B element binding factor, BBF, and repressors regulate the human alpha 1(III) collagen gene (COL3A1).

Type III collagen is found in fetal skin and blood vessels. Previously, we characterized the proximal promoter of the human alpha1(III) collagen gene (COL3A1) using the human rhabdomyosarcoma cell line, A204, and NIH3T3 cells (Yoshino et al., Biochim Biophys Acta, 2005). In the present study, we further analyzed this promoter using additional cell lines, namely a human embryonal rhabdomyosarcoma cell line (RD) and bovine vascular smooth muscle cells (vSMCs), both of which show high expression of type III collagen. Using a luciferase assay, electrophoretic mobility shift assays (EMSA), and DNase footprinting assay, 2 types of multifactor complexes were shown to bind to the DNA region in the vicinity of the B element (- 80 to - 58), depending on the cell type. Next, we used cells stably transfected with a GFP-linked type III collagen promoter fragment for analysis of promoter expression. Usually, transfected cells retained the characteristics of the original cells. However, in several clones derived from RD cells, promoter expression as well as cell shape changed to patterns characteristic of the A204 cell line. Nuclear factors expressed by these clones were also characteristic of the A204 line.</p

CREB-AP1 Protein Complexes Regulate Transcription of the Collagen XXIV Gene (Col24a1) in Osteoblasts

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Increased Cholera Toxin-, and Forskolin-induced Cyclic AMP Accumulations in Psoriatic Involved Versus Uninvolved or Normal Human Epidermis

Psoriatic involved epidermis reveals variously altered receptor-adenylate cyclase responses; among them the most prominent is defective beta-adrenergic adenylate cyclase response, which is normally the major receptor-adenylate cyclase system of human epidermis. It is known that activation of hormone-stimulated adenylate cyclase, a membrane-bound enzyme complex, requires functional coupling of at least 3 distinct subunits: 1) receptor subunit (R), 2) guanine nucleotide binding protein (G), and 3) catalytic subunit (C). The precise nature of the beta-adrenergic defect in the psoriatic epidermis, however, remains to be determined, especially in terms of G and C function. Using the involved and uninvolved skin from psoriatic patients, we investigated effects of cholera toxin (which monitors G-C interaction) and forskolin (which monitors C function) on the adenylate cyclase system of epidermis, which were compared with those of normal human epidermis. Both agents increased cyclic AMP levels of involved, uninvolved, and normal human epidermis. Marked accumulations were observed in the presence of cyclic nucleotide phosphodiesterase inhibitor, isobutyl- methyixanthine (IBMX); without the phosphodiesterase inhibitor, the effect of each agent was minimal. Comparison of the effects of cholera toxin revealed that the psoriatic involved epidermis accumulates much more cyclic AMP than the uninvolved epidermis (involved: 193 ± 65; uninvolved: 117 ± 54 pmoles/mg protein/5 h). Similarly forskolin-induced cyclic AMP accumulations of the involved epidermis were much more than those of uninvolved epidermis (involved: 374 ± 152; uninvolved: 101 ± 41 pmoles/mg protein/2 h). Those of normal human epidermis were not significantly different from those of uninvolved epidermis (cholera toxin: 99 ± 36 pmoles/mg protein/5 h; forskolin: 84 ± 22 pmoles/mg protein/2 h). Our results indicate that G and C function and their interaction is not defective (but rather increased) in the posoriatic involved epidermis. This suggests that the defective beta-adrenergic response of psoriatic involved epidermis reflects defective R or R-G interaction of the epidermal adenylate cyclase system

円盤投げ動作における身体重心速度が円盤速度と円盤+投擲者角運動量に及ぼす効果

The purpose of this study is to analyze the relationships for a discus thrower between : a) the linear velocity of the body center of mass of a discus thrower and discus linear velocity at the moment of release, b) the linear velocity of the body center of mass of a discus thrower and the angular momentum of the discus-and-thrower-system in relation to its vertical axis during discus throwing. The subjects of this study were ten (right-handed) male track and field athletes. Two high speed video cameras operating at 250 fields per second were used to obtain three-dimensional data on the performances of the subjects for each trial. The trial in which each subject recorded his best throw in the test was selected for analysis. A significant correlation (r=0.699, p<0.05) was found between the linear velocity of the body center of mass of a discus thrower with his left foot on the ground (L-on) and discus linear velocity at the moment of release (Rel). The linear velocity of the body center of mass of a discus thrower and the angular momentum of the discus-and-thrower-system in relation to its vertical axis were not correlated significantly with each other during discus throwing. These data provide evidence that the linear velocity of the body center of mass of a discus thrower should be increased as much as possible towards the delivery phase (DV) in order to obtain optimum discus linear velocity at the moment of release (Rel) during discus throwing