Regulation of cell adhesion by protein-tyrosine phosphatases. I. Cell-matrix adhesion

Protein-tyrosine phosphatases are key regulators of protein tyrosine phosphorylation. More than merely terminating the pathways initiated by protein-tyrosine kinases, phosphatases are active participants in many signaling pathways. Signals involving tyrosine phosphorylation are frequently generated in response to cell-matrix adhesion. In addition, high levels of protein tyrosine phosphorylation generally promote disassembly or turnover of adhesions. In this brief review, we will discuss the role of protein-tyrosine phosphatases in cell-matrix adhesions.     

PTP-PEST couples membrane protrusion and tail retraction via VAV2 and p190RhoGAP

Cell motility is regulated by a balance between forward protrusion and tail retraction. These phenomena are controlled by a spatial asymmetry in signals at the front and the back of the cell. We show here that the protein-tyrosine phosphatase, PTP-PEST, is required for the coupling of protrusion and retraction during cell migration. PTP-PEST null fibroblasts, which are blocked in migration, exhibit exaggerated protrusions at the leading edge and long, unretracted tails in the rear. This altered morphology is accompanied by changes in the activity of Rho GTPases, Rac1 and RhoA, which mediate protrusion and retraction, respectively. PTP-PEST null cells exhibit enhanced Rac1 activity and decreased RhoA activity. We further show that PTP-PEST directly targets the upstream regulators of Rac1 and RhoA, VAV2 and p190RhoGAP. Moreover, we demonstrate that the activities of VAV2 and p190RhoGAP are regulated by PTP-PEST. Finally, we present evidence indicating the VAV2 can be regulated by integrin-mediated adhesion. These data suggest that PTP-PEST couples protrusion and retraction by acting on VAV2 and p190RhoGAP to reciprocally modulate the activity of Rac1 and RhoA.     

Molecular characterization of beta-lactamase genes blaA and blaB of Yersinia enterocolitica biovar 1A

The beta-lactamase genes blaA and blaB were detected by PCR amplification in strains of Yersinia enterocolitica biovar 1A isolated from India, Germany, France and the USA. Both genes were detected in all strains. Polymerase chain reaction-restriction fragment length polymorphism revealed genetic heterogeneity in blaA but not in blaB. Cluster analysis of blaA restriction profiles grouped the strains into three groups. The blaA gene of Y. enterocolitica biovar 1A showed a high degree of sequence homology to that of Y. enterocolitica 8081 (biovar 1B) and Y. enterocolitica Y-56 (biovar 4), whereas homology was low with class A beta-lactamase genes of other members of the family Enterobacteriaceae. The pI 8.7 of enzyme Bla-A of Y. enterocolitica biovar 1A was similar to that of biovars 2, 3 and 4. The enzyme Bla-B focused at 6.8 and 7.1, indicating that biovar 1A strains produced a 'B-like' enzyme. This is the first study to have investigated the genetic heterogeneity of the beta-lactamase genes of Y. enterocolitica.     

Antioxidant response in sesame plants grown on industrially contaminated soil: effect on oil yield and tolerance to lipid peroxidation

The plants of sesame white (Sesamum indicum L. var. T55) grown on tannery sludge (TS) contaminated soil have shown that Cr level in the seeds was found below detection limits in 10% and 25% TS, however, the levels of Ni, Pb and Cd were found above the recommended limits. In roots, the level of antioxidants increased in the plants grown upto 35% TS at 30d over their respective controls. Total chlorophyll content increased significantly (p<0.5) in the plants (leaves) grown on lower sludge amendments (upto 35% TS at 30d and 25% TS at 60d) over their respective controls. In addition, the oil content increased (35% increase over control) in the plants grown on 35% TS. No significant change was observed in thiobarbituric acid reactive substances (TBARS), a lipid peroxidation index, in the plants (upto 50% TS). The number of trichomes in the leaves of treated plants was found more than control. In lower and upper leaves surfaces, the anterior end of the trichomes was found acute tipped and bent downwards, whereas, the trichome tip was straight and blunt in control. The stomata on upper and lower surfaces of the leaves were found partially or totally closed in the plants grown on 100% TS as compared to control. The toxicity was observed at higher amendments which are evident from the observed morphological changes and decrease in chlorophyll content. This study concludes that it is not advisable to grow the plants on contaminated area, besides its healthy growth.     

Plant regeneration from alginate-encapsulated shoot tips of Spilanthes acmella (L.) Murr., a medicinally important and herbal pesticidal plant species

This article demonstrates the plantlet regeneration from alginate-encapsulated shoot tips of Spilanthes acmella. Shoot tip explants excised from in vitro proliferated shoots were encapsulated in calcium alginate beads. The best gel complexation for encapsulation of shoot tips was achieved using 3% sodium alginate and 100 mM calcium chloride. Maximum percent response for the conversion of encapsulated shoot tips into plantlets was obtained on growth regulator-free full-strength liquid MS (Murashige and Skoog, Physiol Plant 15:473–497, 1962) medium. The addition of MS nutrients in alginate matrix was found to have pronounced effect on shoot and root emergence from alginate beads. Encapsulated shoot tips could be stored at low temperature (4°C) up to 60 days. Plantlets regenerated from encapsulated shoot tips were acclimatized successfully. The present synthetic seed technology could be useful in large-scale propagation as well as short-term conservation and germplasm distribution and exchange of Spilanthes acmella. S. K. Singh and M. K. Rai contributed equally to this work.     

Sex‐specific effects of gestational and lactational coexposure to lead and cadmium on hepatic phase I and phase II xenobiotic/steroid‐metabolizing enzymes and antioxidant status

Liver has evolved complex enzymatic mechanisms to detoxify a wide array of xenobiotic substances, ranging from dietary components to environmental toxins to pharmaceuticals. Activities of many steroid‐metabolizing enzymes in adult rat liver microsomes are sexually differentiated. Toxic effects of lead and cadmium on hepatic tissue have been well established in our earlier studies. We thus monitored the effects of gestational and lactational coexposure to lead and cadmium on hepatic phase I and phase II xenobiotic‐ and steroid‐metabolizing enzyme activities in both male and female F1 generation postnatal day (PND) 56 rats. Adult pregnant female rats were treated subcutaneously [0.05 mg/(kg body wt. day)] with sodium acetate (control group), lead acetate, and cadmium acetate separately and in combination throughout the gestational and lactational period. Hepatic phase I xenobiotic‐metabolizing enzymes (NADPH‐ and NADH‐cytochrome c reductase) activities significantly decreased significantly in all the metal‐treated groups in both PND 56 male and female rats as compared with the control group. Hepatic phase II enzymes (uridine diphosphate‐glucuronosyl transferase, γ‐glutamyl transpeptidase, glutathione‐S‐transferase, 17‐β‐hydroxysteroid oxidoreductase) were also highly susceptible to all the metal‐treated groups. The observed alterations in the oxidative stress and biochemical parameters in the liver of F1 generation male and female rats resulted from an independent effect of lead and/or cadmium and also from their interaction. Results suggest that early developmental exposure to lead and cadmium both alone and in combination can suppress the hepatic xenobiotic‐metabolizing enzyme activities in the liver of F1 generation male and female rats in a sex‐dependent manner. © 2009 Wiley Periodicals, Inc. J Biochem Mol Toxicol 23:419–431, 2009; Published online in Wiley InterScience ( www.interscience.wiley.com ). DOI 10.1002/jbt.20305     

Occurrence of Macrosiphum hellebori Theobald & Walton (Hemiptera: Aphididae) in Australia

The European aphid species, Macrosiphum hellebori , the hellebore aphid, is recorded from Australia, having been found in Melbourne, Victoria and in Canberra, Australian Capital Territory, colonising ornamental hellebores, Helleborus species (Ranunculaceae). Insect associates of M. hellebori are recorded, including the hymenopterous primary parasite Lysiphlebus testaceipes (Cresson) (Braconidae: Aphidiinae). Keys are provided for the identification of the three species of Macrosiphum Passerini known from Australia.     

Growth,metal accumulation and yield performance of Brassica campestris L. (cv. Pusa Jaikisan) grown on soil amended with tannery sludge/fly ash mixture

Application of sewage sludge and fly ash (FA) to soil is gaining practice and has become an alternative to chemical fertilizers in many countries. The present study aimed to assess the potential of fly ash–tannery sludge (TS) mixture in two ratios {4 (TS):1 (FA), denoted as A and 4 (TS):2 (FA) denoted as B} and different amendments were made with soil to study the yield and metal availability and their uptake by Brassica campestris L. (cv. Pusa Jaikisan). Experimental data showed that increase in dry matter yield and oil content of B. campestris was noted in lower amendments of soil with mixture A. However, total chlorophyll content of the leaf increased significantly, whereas carotenoid content showed non-significant increase in all the amendments with mixtures A and B as compared to control. The translocation of most of the tested metals (Pb, Mn, Cd, Ni, and Fe) in the shoot of the plant was found higher except Cr, Cu, and Co. Overall, experimental results showed that mixture A (≥10%) was found to be suitable due to better yield and less accumulation of metal in the seed of B. campestris.     

Does post-fire abiotic habitat filtering create divergent plant communities in black spruce forests of eastern Canada?

We investigated the role of post-fire residual organic matter (ROM) thickness as a driver of community assembly in eastern Newfoundland. We hypothesized that if post-fire community assembly is predominantly controlled by ROM thickness (an abiotic habitat filter), then post-fire species composition and functional traits should correspond to the depth and distribution of ROM. However, if species interactions (biotic filter) are the primary constraints on community assembly, then post-fire species composition and their functional traits should be independent of the depth and distribution of ROM. We tested these predictions in three relatively mature plant communities, Kalmia angustifolia heath, black spruce (Picea mariana)-Kalmia shrub savannah and black spruce forest. Through pre-fire stand reconstruction, we found evidence that the three communities originated from black spruce forest. ROM thickness in heath was almost twice that of shrub savannah and six times more than forest, suggesting a gradient in fire severity. Distribution of ROM corresponded to patterns in vegetation dominance, where thick ROM (>2 cm) filtered out black spruce in favour of Kalmia. ROM thickness was a strong predictor of vegetation composition and function between heath and forest, but this was not found between the shrub savannah and forest. We attribute this to species interactions and allelopathy, which may have become important when ROM thickness was suitable for both seed (black spruce) and vegetative (Kalmia) regenerating species. Thus, priority effects or “who came first” may have lead to shrub savannah formation when ROM thickness was ~2 cm. We conclude that abiotic habitat filtering of thick ROM (>2 cm) on (primarily) species’ regeneration traits was the primary driver of community divergence from forest to heath and shrub savannah.     

Acidophilic tannase from marine Aspergillus awamori BTMFW032

Aspergillus awamori BTMFW032, isolated from sea water, produced tannase as extracellular enzyme under submerged culture conditions. Enzyme with a specific activity of 2761.89 IU/mg protein, a final yield of 0.51 %, and a purification fold of 6.32 was obtained after purification to homogeneity by ultrafiltration and gel filtration. SDS-PAGE analyses under non- reducing and reducing conditions yielded a single band of 230 kDa and 37.8 kDa, respectively, indicating presence of six identical monomers. pI of 4.4 and 8.02 % carbohydrate content in the enzyme were observed. Optimal temperature was 30oC, although the enzyme was active at 5-80 oC. Two pH optima, pH 2 and pH 8, were recorded and the enzyme was stable only at pH 2.0 for 24 h. Methylgallate recorded maximal affinity and K(m) and V(max) were recorded, respectively, as 1.9 X 10?3 M and 830 micronmol/min. Impact of several metal salts, solvents, surfactants, and typical enzyme inhibitors on tannase activity were determined to establish the novelty of the enzyme. Gene encoding tannase isolated from A. awamori is 1.232 kb and nucleic acid sequence analysis revealed an open reading frame consisting of 1122 bp (374 amino acids) of one stretch in -1 strand. In-silico analyses of gene sequences and comparison with reported sequences of other species of Aspergillus indicated that the acidophilic tannase from marine A. awamori is differs from that of other reported species.