Synthesis,degradation, and subcellular localization of proteins encoded by the primary response genes TIS7/PC4 and TIS21/PC3

Murine TIS7 and TIS21 cDNAs were cloned from phorbol ester-induced Swiss 3T3 cells. The cognate rat cDNAs. PC4 and PC3, were cloned from nerve growth factor (NGF)-treated PC12 pheochromocytoma cells. The TIS7/PC4 and TIS21/PC3 primary response genes are rapidly and transiently induced in response to serum, phorbol esters, and polypeptide growth factors in quiescent Swiss 3T3 cells and by NGF and other ligands in PC12 cells. In both 3T3 and PC12 cells the appearance of the TIS21/PC3 message precedes that of TIS7/PC4 message following ligand stimulation, suggesting that the TIS21/PC3 protein is likely to be synthesized more rapidly than the TIS7/PC4 protein. Using antisera prepared against recombinant TIS21 and TIS7 proteins, we find that the TIS21/PC3 protein is, indeed, synthesized more rapidly than the TIS7/PC4 protein following stimulation in both 3T3 and PC12 cells. In addition, “pulse-chase” experiments demonstrate that the TIS21/PC3 protein is degraded much more rapidly than the TIS7/PC4 protein. The sequences of the predicted PC3 and PC4 proteins have lead to the speculation that these two proteins may both be secreted from cells following stimulation. The PC4 protein is reported to have some sequence similarity to interferons. The TIS21/PC3 protein contains a presumptive leader sequence. Using our antisera to the recombinant proteins, however, we cannot detect secretion of radiolabelled TIS7/PC4 or TIS21/PC3 protein. Immunohistochemical and subcellular fractionation experiments suggest that the TIS7 protein is a membrane associated, non-nuclear intracellular protein. The TIS21 protein, in contrast, is' a non-nuclear, soluble intracellular protein. © 1994 Wiley-Liss, Inc.     

A new alpha-helical coiled coil protein encoded by the Salmonella typhimurium virulence plasmid.

A new protein of Salmonella typhimurium was identified and characterized. The gene (tlpA) encoding this protein (TlpA) was isolated from the large virulence-associated plasmid of S. typhimurium and sequenced in order to predict the primary structure of TlpA. tlpA encodes a 371-amino acid soluble protein with a calculated M(r) of 41600 and pI of 4.63. Secondary structure predictions and sequence statistics of TlpA indicated a predominant alpha-helical configuration and presence of heptapeptide repeat motifs characteristic of coiled coil proteins. Purified TlpA was shown to have biochemical properties similar to those of coiled coil proteins, including adoption of an alpha-helical configuration and a tendency to form homodimers. Furthermore, TlpA possessed heat resistance, evidence for a chain register and altered mobility in urea/sodium dodecyl sulfate-polyacrylamide gel electrophoresis gels which are characteristics of tropomyosins. TlpA shows 32% overall sequence similarity with rat cardiac myosin and 36% similarity with horse platelet beta-tropomyosin over 226 residues, whereas selected regions possessed significant sequence identities with myosins, tropomyosins, and alpha-helical surface proteins of Streptococcus pyogenes. Our results indicate that TlpA represents a new member of prokaryotic coiled coil proteins.     

Modulation of sodium-sensitive GTPase by partial opiate agonists. An explanation for the dual requirement for Na+ and GTP in inhibitory regulation of adenylate cyclase

Opiates and opioid peptides inhibit adenylate cyclase and stimulate specific low Km GTPase activity in membranes from neuroblastoma x glioma NG108-15 hybrid cells. The effects of opiate agonists on both enzymes are mediated by high affinity stereospecific receptors and require Mg2+, GTP, and Na+. In the presence of Mg2+, Na+ inhibits basal GTPase activity; opiates stimulate GTP hydrolysis by antagonizing the Na+-induced inhibition. Activation of GTPase leads, in turn, to inactivation of GTP-stimulated adenylate cyclase activity. The intrinsic activities (or efficacies) of a series of opiates are identical for stimulation of GTPase and inhibition of adenylate cyclase. These results provide a mechanism for the dual requirement for Na+ and GTP in the inhibitory coupling of opiate receptors to the adenylate cyclase system in these cells and may be of general significance to the action of other inhibitory hormones.     

Considerations in the grouping of plant and fungal taxa for an epidemiologic study

Although exposure to airborne pollen grains and fungal spores has been implicated as a causative factor for acute exacerbation of asthma, the few epidemiologic studies that have attempted to evaluate the relationship between these bioaerosols and asthma have used only total counts (ignoring the relative importance of different taxa) or a few predominant pollen or spore types (ignoring less abundant but potentially relevant groups). This paper reports the development of hypothesis‐driven exposure metrics (based on known aeroallergen associations with allergic asthma and other hypersensitivity diseases, pollen allergen cross‐reactivity, and the presence of local sources in the city of Fresno, California, USA) for a 3.5 year epidemiologic study of childhood asthma. Outdoor regional and neighborhood concentrations of pollen and spores were measured using Hirst‐type, 7‐day samplers. Indoor and outdoor residential concentrations were measured at 84 selected homes with similar 24‐hour slit impactors. All pollen and spore concentrations were recorded in 2‐hour intervals to assist in understanding diurnal fluctuations in aeroallergen concentrations, identify exposures during the time periods that children are outdoors, and study interaction between aeroallergens and other air contaminants, which were the primary focus of the study. The 124 pollen taxa that were observed were reduced to 15 categories and the 66 fungal and algal taxa were reduced to five categories that will be used in microenvironmental models to generate individual daily exposure estimates for each of the 315 children. These new exposure metrics will allow examination of health effects for taxa traditionally associated with allergy and those with locally elevated concentrations in combination with exposures to other indoor and outdoor air contaminants.     

Random Partition Models and Exchangeability for Bayesian Identification of Population Structure

We introduce a Bayesian theoretical formulation of the statistical learning problem concerning the genetic structure of populations. The two key concepts in our derivation are exchangeability in its various forms and random allocation models. Implications of our results to empirical investigation of the population structure are discussed.     

The fate of discarded appendicularian houses: degradation by the copepod, Microsetella norvegica, and other agents

Despite the potential importance of zooplankton in degradationof marine snow, the association of colonising zooplankton withdiscarded appendicularian houses has not been investigated innorthern areas. We sampled the vertical distributions of appendicularians,houses and potential zooplankton colonisers at two stationsin the central North Sea during late summer. In addition, grazingexperiments were performed with the copepod Microsetella norvegica,which was assumed to be the main contributor to house degradation.The results were used in (i) inverse modelling, to estimatethe factors which were significant in shaping the vertical distributionof houses and (ii) calculations to estimate potential housedegradation rates. M. Norvegica was able to feed on appendicularianhouses, with feeding rates up to 0.42 g C (g C)^–1 day^–1(0.14 µg C ind.^–1 day^–1). The model resultssuggested that the vertical distribution of houses was shapedby sinking of houses, bacterial degradation and feeding of M.norvegica and invertebrate larvae. The estimated community degradationrate by M. norvegica was low, whereas invertebrate larvae haddegradation rates close to bacterial degradation. We concludethat at the typical concentrations of M. norvegica in the NorthSea (10⁴ ind. m^–2), its role in marine snow degradationis likely to be small. Degradation by other zooplankton groups,such as invertebrate larvae, can, however, be substantial.