Platelet-derived growth factor-specific regulation of the JE promoter in rat aortic smooth muscle cells

JE is a member of the family of "immediate early" genes induced by growth factors and cytokines. JE encodes a low molecular weight secretory glycoprotein analogous to the human monocyte chemoattractant protein, MCP-1. JE and MCP-1 proteins are thought to play an important role in inflammation and in the recruitment of monocyte/macrophages to the vessel wall during the development of atherosclerosis. We have previously reported that the induction of JE in rat aortic smooth muscle cells (SMC) was specific to platelet-derived growth factor (PDGF) and was not seen with other growth agonists. Using a luciferase reporter system and transient transfection assays of rat aortic SMC, we now report the identification of a region in the proximal rat JE promoter that is responsive to PDGF but not to other growth factors (angiotensin II and alpha-thrombin) or cytokines (interleukin 1-beta and tumor necrosis factor-alpha). The full response to PDGF (approximately 6-fold) requires the cooperative activity of two potentially novel cis-acting elements, at positions -146 to -128 and -84 to -59. While each element produces a different pattern in electrophoretic mobility shift assays, they appear to bind the same PDGF-responsive species. Further analysis of these regions should provide important insights into PDGF-specific responses in vascular SMC.     

Assembly and analysis of conical models for the HIV-1 core

The genome of the human immunodeficiency virus (HIV) is packaged within an unusual conical core particle located at the center of the infectious virion. The core is composed of a complex of the NC (nucleocapsid) protein and genomic RNA, surrounded by a shell of the CA (capsid) protein. A method was developed for assembling cones in vitro using pure recombinant HIV-1 CA-NC fusion proteins and RNA templates. These synthetic cores are capped at both ends and appear similar in size and morphology to authentic viral cores. It is proposed that both viral and synthetic cores are organized on conical hexagonal lattices, which by Euler's theorem requires quantization of their cone angles. Electron microscopic analyses revealed that the cone angles of synthetic cores were indeed quantized into the five allowed angles. The viral core and most synthetic cones exhibited cone angles of approximately 19 degrees (the narrowest of the allowed angles). These observations suggest that the core of HIV is organized on the principles of a fullerene cone, in analogy to structures recently observed for elemental carbon.     

Activation of transducin guanosine triphosphatase by two proteins of the RGS family

RGS proteins (regulators of G protein signaling) constitute a newly appreciated group of negative regulators of G protein signaling. Several members of this group stimulate the guanosine triphosphatase (GTPase) activity of various G protein alpha-subunits, including the photoreceptor G protein, transducin. In photoreceptor cells transducin GTPase is known to be substantially accelerated by the coordinated action of the gamma-subunit of its effector enzyme, cGMP phosphodiesterase (PDE gamma), and another yet unidentified membrane-associated protein factor. Here we test the possibility that this factor belongs to the RGS family of GTPase stimulators. We report a detailed kinetic analysis of transducin GTPase activation by two members of the RGS family, RGS4 and G alpha interacting protein (GAIP). RGS4, being at least 5-fold more potent than GAIP, stimulates the rate of transducin GTPase by 2 orders of magnitude. Neither RGS4 nor GAIP requires PDE gamma for activating transducin. Rather, PDE gamma causes a partial reversal of transducin GTPase activation by RGS proteins. The effect of PDE gamma is based on a decreased apparent affinity of RGS for the alpha-subunit of transducin. Our observations indicate that GTPase activity of transducin can be activated by at least two distinct mechanisms, one based on the action of RGS proteins alone and another involving the cooperative action of the effector enzyme and another protein.     

Unique reactive site domains of neuroendocrine isoforms of alpha 1-antichymotrypsin from bovine adrenal medulla and pituitary revealed by molecular cloning

Molecular cloning of bovine adrenal medulla (AM) and pituitary (Pit) alpha 1-antichymotrypsin cDNAs indicated novel isoforms of ACT. The deduced primary sequences indicated that the AM ACT and Pit ACT possess COOH-terminal reactive-site domains that are characteristic of serpins (serine protease inhibitors). Of high interest was the finding of unique reactive sites within AM ACT and Pit ACT which are predicted to possess Arg as P1 residue. Arginine as P1 residue parallels the cleavage specificity of neuroendocrine prohormone processing enzymes cleaving at basic residues. Furthermore, RT-PCR indicated tissue-specific expression of AM and Pit ACT mRNAs. The AM and Pit isoforms of ACT may regulate novel target proteases involved in neuroendocrine function.     

The effectiveness of a tobacco prevention program with adolescents living in a tobacco-producing region

OBJECTIVES: This study investigated the efficacy of a social-influences tobacco prevention program conducted with adolescents living in a high tobacco production area. METHODS: Students in 10 experimental schools completed the tobacco prevention program and a booster intervention. Control students received health education as usual. RESULTS: After 2 years of treatment, smoking rates in the treatment group (vs the control group) were lower for 30-day, 7-day, and 24-hour smoking. The intervention had more of an impact on those who were involved in raising tobacco than it did on those not involved in raising tobacco. CONCLUSIONS: Although modest, effects were achieved with minimal intervention time in a high-risk group, indicating that social-influences prevention programs may be effective in such groups.