Physiological Effects of a and {alpha} Sexual Agglutination Substances on Mating Reaction in the Yeast Saccharomyces cerevisiae

The sex-specific glycoprotein agglutination substance, responsiblefor sexual agglutination, solubilized from the surface of haploidcells of a or a mating type by the autoclave method had thefollowing effects on mating reaction in Saccharomyces cerevisiae.Sexual agglutination was inhibited by the agglutination substanceof the opposite mating type in living cells as well as in heat-killedcells. Formation of zygotes was completely inhibited, when botha and a cells were treated with the agglutination substanceof the opposite mating type. The a and a agglutination substanceswere inactivated by cells of the opposite mating type, withthe degree of inactivation being greater for the former. Theenzyme responsible for the inactivation of a agglutination substanceseems to be carboxypeptidase Y. ¹ This paper is dedicated to the late Professor J. Ashida, KyotoUniversity. ² Present address: Department of Plant Pathology, Universityof California, Davis, CA. 95616, U.S.A. (Received November 1, 1982; Accepted January 19, 1983)     

Difference in growth factor requirements of rat 3Y1 cells among growth in mass culture, clonal growth in low density culture, and stimulation to enter S phase in resting culture

A semiserum-free medium was developed for monolayer culture of rat 3Y1 fibroblastic cells. The main components of the developed medium added to Dulbecco's modified Eagle's medium (DMEM) were insulin, transferrin, epidermal growth factor, poly-D-lysine, bovine albumin, oleic acid, and bovine alpha-globulin. In this medium, 3Y1 cells grew in mass culture at much the same rate as in DMEM supplemented with 10% fetal bovine serum (FBS), and colonies, albeit of smaller sizes, did form. Virally transformed derivatives of 3Y1 (simian virus 40-3Y1, polyoma virus-3Y1 and adenovirus type 12-3Y1) also formed colonies in the semiserum-free medium. When trypsinized 3Y1 cells were seeded with the medium lacking alpha-globulin, neither growth in the mass culture nor clonal growth in the low density culture (clonal growth) occurred. In this case, cell spreading was inhibited by albumin, and this inhibition was overcome by adding alpha-globulin or treating dishes with serum. When albumin was excluded from the semiserum-free medium, clonal growth did not occur, whereas growth in mass culture and stimulation of DNA synthesis in the resting mass culture (stimulation of DNA synthesis) were not so drastically affected. When oleic acid was removed, growth in mass culture was inhibited considerably, but no considerable effect was seen on clonal growth or on stimulation of DNA synthesis. In the absence of insulin, stimulation of DNA synthesis was inhibited more markedly than when other components were removed, but such was not the case with growth in mass culture and clonal growth.     

Fluorescence polarization as a tool to study lectin-sugar interaction

The binding ofRicinus communis agglutinin andAbrus agglutinin to 4-methylumbelliferyl β-D-galactopyranoside was studied by equilibrium dialysis, fluo-rescence quenching and fluorescence polarization. The number of binding sites and the association constant value obtained by fluorescence polarization for bothRicinus communis agglutinin andAbrus agglutinin are in close agreement with those obtained by the other methods. This indicates the potential of ligand-fluorescence polarization measurements in the investigation of lectin-sugar interactions.     

Lectins from<Emphasis Type="Italic">Griffonia simplicifolia</Emphasis> seeds

The physical-chemical and carbohydrate binding specificity ofGriffonia simplicifolia I (GS I) isolectins, one of the 4 lectins isolated fromGriffonia simplicifolia seeds, are described.Association constants for the binding of methyl α- and β-D-galactopyranoside and methyl 2-acetamido-2-deoxy-α-D-galactopyranoside to the A₄, A₂ B₂ and B₄ isolectins are reported.Precipitation reactions of theGriffonia simplicifolia isolectins with guaran and type B blood group substance are described.The hypothesis that subunit B is a precursor of subunit A, a process involving proteolytic cleavage of the B subunit, was tested by conducting structural studies on the 2 subunits. The results indicated that the A and B subunits are probably products of 2 separate but closely related, possibly contiguous genes.     

Haemolysis of rabbit erythrocytes by a lectin from the seeds of<Emphasis Type="Italic">Croton tiglium</Emphasis>

The kinetics of haemolysis of rabbit erythrocytes byCroton tiglium lectin was studied as a function of concentration of the lectin and erythrocytes. The length of the prelytic period decreased with increasing lectin concentrations, indicating that the secondary events at the membrane which follow the binding of the lectin to cell surface carbohydrate receptors are accelerated at higher surface concentrations of the lectin. The rate or extent of haemolysis was not affected by the inclusion of ions like K⁺, Ca²⁺ and Mg²⁺ in the medium or by the substitution of ionic medium by a non-ionic medium. The inhibition of haemagglutination and haemolysis of rabbit red cells byCroton tiglium lectin by antilectin rabbit serum was observed. A possible mechanism of haemolysis by the lectin is discussed.     

Plant lectins specific for N-acetyl-β-D-galactosamine

N-Acetyl-D-galactosamine in β-linkage being ubiquitous in cell surface glycoproteins, their interaction with lectins specific for this sugar moiety may be a significant event in cell adhesion phenomena. This article discusses the common β-N-acetyl galactosamine-specific lectins, with particular stress on the lectin from winged beans (Psophocarpus tetragonolobus).     

The ultrastructural basis of the permeability of arterial endothelium to horseradish peroxidase

The thoracic aorta and basilar artery, in which the incidence of atherosclerosis is known to be different, were examined to elucidate the correlation between the structure of the intercellular cleft junction between adjacent endothelial cells and its permeability to HRP. Tannic acid or HRP in the vessel lumen passed through the intercellular clefts of the thoracic aorta into the subendothelial space, whereas in the basilar artery they were unable to penetrate beyond the tight junction of the intercellular clefts. Freeze-fracture replicas revealed that the tight junctions of the thoracic aorta consisted of one to two junctional strands in most areas of the cleaved planes, with discontinuities in some places, whereas those of the basilar artery consisted of a continuous belt-like meshwork of six anastomosing junctional strands on average. These observations confirm that the structure of endothelial junctions in arteries has a close correlation with the permeability of the intercellular clefts to HRP.     

Role of Peroxidase when Hydroxyproline-rich Protein in Plant Cell Walls is increased by Ethylene

Ethylene may control the growth of plant cells by regulating hydroxylation of specific wall proteins.     

Genetic Control of Variable and Constant Regions of Immunoglobulin Heavy Chains

IMMUNOGLOBULIN polypeptide chains consist of two well defined regions designated the “variable region” and the “constant region”. Whereas great diversity exists in amino-acid sequences of variable regions, the constant regions of a given subclass of heavy chains (C_H)^* are essentially invariant in sequence^{1, 2}. Exceptions are the allelic forms, such as the rabbit allotypes A14 and A15^{3, 4}, where a threonine-alanine interchange occurs in the constant region of γ chains (Appella, Chersi, R. G. M. and Dubiski, in preparation). The markers unique to a chains (for example, A14-A15) are closely linked to allotypic markers at the a locus (a1, a2, a3)^{3, 4} which seem to be present on four different Ig heavy chain classes (α, γ, ε, µ)^5–7. These puzzling observations can be explained if the a locus determinants are variable region markers which reflect genetically controlled differences in some relatively constant residues within the V_H region sequences⁷.     

Mitochondrial Ribosomes in HeLa Cells

HeLa cell mitochondria contain 60S ribosomes which seem to consist of subunits of 45S and 35S particles. The 16S and 12S RNA components are coded by mitochondrial DNA.