Interaction between the Msh2 and Msh6 Nucleotide-binding Sites in the Saccharomyces cerevisiae Msh2-Msh6 Complex

Indirect evidence has suggested that the Msh2-Msh6 mispair-binding complex undergoes conformational changes upon binding of ATP and mispairs, resulting in the formation of Msh2-Msh6 sliding clamps and licensing the formation of Msh2-Msh6-Mlh1-Pms1 ternary complexes. Here, we have studied eight mutant Msh2-Msh6 complexes with defective responses to nucleotide binding and/or mispair binding and used them to study the conformational changes required for sliding clamp formation and ternary complex assembly. ATP binding to the Msh6 nucleotide-binding site results in a conformational change that allows binding of ATP to the Msh2 nucleotide-binding site, although ATP binding to the two nucleotide-binding sites appears to be uncoupled in some mutant complexes. The formation of Msh2-Msh6-Mlh1-Pms1 ternary complexes requires ATP binding to only the Msh6 nucleotide-binding site, whereas the formation of Msh2-Msh6 sliding clamps requires ATP binding to both the Msh2 and Msh6 nucleotide-binding sites. In addition, the properties of the different mutant complexes suggest that distinct conformational states mediated by communication between the Msh2 and Msh6 nucleotide-binding sites are required for the formation of ternary complexes and sliding clamps.     

Gradients in maize roots: local elongation and pH

The elongation rate, the gradient of the local elongation rate and the surface pH of maize roots were measured over 12 h. A data bank was constituted by storing these values. By sorting these results on the basis of different elongation rates, different classes of root were obtained. Two classes were chosen: the low-growth roots and the high-growth roots. The mean growth of these two root classes was stable with time and differed significantly from one another. The surface pH of the elongation zone was the same for the roots of these two classes, but the roots selected for their higher growth rate had a larger acid efflux in this zone.     

The Effect of Leaf Age on Gas Exchange and Malate Accumulation in C3-CAM Plant Marrubium Frivaldszkyanum (Lamiaceae)

For the first time the expression of C₃ and CAM in the leaves of different age of Marrubium frivaldszkyanum Boiss, is reported. With increasing leaf age a typical C₃ photosynthesis pattern and high transpiration rate were found. In older leaves a shift to CAM occurred and the 24-h transpiration water loss decreased. A correlation was established between leaf area and accumulation of malate. Water loss at early stages of leaf expansion may be connected with the shift to CAM and the water economy of the whole plant.     

Ultrastructural studies of endocrine-like cells in the fundic gastric mucosa of the bullfrog,Rana catesbeiana

Summary This study is concerned with electron-microscopic observations on endocrine or paracrine cells in the fundic gastric mucosa of the bullfrog. Also, an attempt was made to identify the histamine-releasing cells involved in the secretagogue response. At least three distinct endocrine-like cell types were found. The classification is based on the appearance of secretory granules and other organelles, and the relationship of endocrine-like cells with other cells in the tissue. The amphibian endocrine-like cells resemble the ECL, D and EC cells of mammals. Type-I (ECL) cells showed degranulation after repeated stimulation with tetragastrin (TG), acetylcholine (ACh) and K⁺ depolarizing solution, all of which release histamine.     

Kinetic Basis for the Conjugation of Auxin by a GH3 Family Indole-acetic Acid-Amido Synthetase

The GH3 family of acyl-acid-amido synthetases catalyze the ATP-dependent formation of amino acid conjugates to modulate levels of active plant hormones, including auxins and jasmonates. Initial biochemical studies of various GH3s show that these enzymes group into three families based on sequence relationships and acyl-acid substrate preference (I, jasmonate-conjugating; II, auxin- and salicylic acid-conjugating; III, benzoate-conjugating); however, little is known about the kinetic and chemical mechanisms of these enzymes. Here we use GH3-8 from Oryza sativa (rice; OsGH3-8), which functions as an indole-acetic acid (IAA)-amido synthetase, for detailed mechanistic studies. Steady-state kinetic analysis shows that the OsGH3-8 requires either Mg²⁺ or Mn²⁺ for maximal activity and is specific for aspartate but accepts asparagine as a substrate with a 45-fold decrease in catalytic efficiency and accepts other auxin analogs, including phenyl-acetic acid, indole butyric acid, and naphthalene-acetic acid, as acyl-acid substrates with 1.4–9-fold reductions in k_cat/K_m relative to IAA. Initial velocity and product inhibition studies indicate that the enzyme uses a Bi Uni Uni Bi Ping Pong reaction sequence. In the first half-reaction, ATP binds first followed by IAA. Next, formation of an adenylated IAA intermediate results in release of pyrophosphate. The second half-reaction begins with binding of aspartate, which reacts with the adenylated intermediate to release IAA-Asp and AMP. Formation of a catalytically competent adenylated-IAA reaction intermediate was confirmed by mass spectrometry. These mechanistic studies provide insight on the reaction catalyzed by the GH3 family of enzymes to modulate plant hormone action.     

The effect of simulated acid rain on performance of the aphid Euceraphis betulae (Koch) on silver birch

Summary The effect of artificial acid rain on the reproduction and survival of the aphid Euceraphis betulae on silver birch was studied in Turku, southern Finland. Eight bioassays were done during 1984–1986. In four of the bioassays the aphids produced 40% to over 100% more progeny on birches watered with dilute sulphuric acid (pH 3.5) than on control trees. In four other cases the performance of aphids did not differ between the treatments. An index of aphid reproduction pooled over the whole study was significantly higher on acid-treated than on control birches. The reproduction of aphids on acid-treated birches was enhanced when precipitation was below long term average, suggesting an interaction between the stress caused by acid treatment and dry periods.     

Activity and distribution of enzymes that interconvert purine bases, ribosides and ribotides in the tomato plant and possible implications for cytokinin metabolism

The activity of the enzymes 5'-nucleotidase (EC 3.1.3.5), adenosine nucleosidase (EC 3.2.2.7), adenine phosphoribosyl transferase (EC 2.4.2.7) and acid phosphatase (EC 3.1.3.2) was determined in sections of tomato plant ( Lycopersicon esculentum Mill. cv. Bellina). The distribution of the enzymes changed markedly during development and a role for these enzymes in cytokinin metabolism is suggested.     

Changes in gill histology of fathead minnows and yellow perch transferred to soft water or acidified soft water with particular reference to chloride cells

Summary Fathead minnows, Pimephales promelas, and yellow perch, Perca flavescens, were transferred from moderately soft Lake Superior water (hardness 45mg/l as CaCO₃) to very soft diluted Lake Superior water (hardness 4.5mg/l). Sulfuric acid was added in some treatments by means of a multichannel diluter. In very soft water, chloride cells proliferated in the gills, especially in the epithelium of the secondary lamellae. When exposed to acid, chloride cells were damaged and less abundant in the secondary lamellae, and blood osmolality was reduced at pH 5.0 (x = 188 mOsm/kg, 9 days exposure; normal 280 mOsm/kg) for the minnows and pH 4.1 (x = 218 mOsm/kg, 58 days exposure; normal 329 mOsm/kg) for the perch. Certain chloride cells which form gland-like clusters in the primary lamellae of perch gills showed little damage even at pH 4.1. The present study supports the view that chloride cells proliferate in very soft fresh water to help maintain ionic balances, and that damage to these cells in acidified soft water may be related to diminished ionoregulatory capacity. The greater acid tolerance of chloride cells of, and the higher blood osmolality maintained by, perch could help to explain the greater tolerance of this species to low pH. In some cases, a species' ability to acclimate to very soft water and acidified soft water may depend upon the number, distribution, and physiology of its chloride cells.