Pre-B cell antigen receptor-mediated signal inhibits CD24-induced apoptosis in human pre-B cells

We previously reported that the cross-linking of cluster of differentiation (CD)24 induces apoptosis in Burkitt's lymphoma cells and that this phenomenon can be enhanced by a B cell Ag receptor (BCR)-mediated signal. In this study, we extend our previous observation and report that CD24 also mediated apoptosis in human precursor-B acute lymphoblastic leukemia cell lines in the pro-B and pre-B stages accompanying activation of multiple caspases. Interestingly, simultaneous cross-linking of pre-BCR clearly inhibited CD24-mediated apoptosis in pre-B cells. We also observed that mitogen-activated protein kinases (MAPKs) were involved in the regulation of this apoptotic process. Pre-BCR cross-linking induced prompt and strong activation of extracellular signal-regulated kinase 1, whereas CD24 cross-linking induced the sustained activation of p38 MAPK, following weak extracellular signal-regulated kinase 1 activation. SC68376, a specific inhibitor of p38 MAPK, inhibited apoptosis induction by CD24 cross-linking, whereas anisomycin, an activator of p38 MAPK, enhanced the apoptosis. In addition, PD98059, a specific inhibitor of MEK-1, enhanced apoptosis induction by CD24 cross-linking and reduced the antiapoptotic effects of pre-BCR cross-linking. Collectively, whether pre-B cells survive or die may be determined by the magnitude of MAPK activation, which is regulated by cell surface molecules. Our findings should be important to understanding the role of CD24-mediated cell signaling in early B cell development.     

Cis1/Atg31 is required for autophagosome formation in Saccharomyces cerevisiae

Autophagy is the bulk degradation of cytosolic materials in lysosomes/vacuoles of eukaryotic cells. In the yeast Saccharomyces cerevisiae, 17 Atg proteins are known to be involved in autophagosome formation. Genome wide analyses have shown that Atg17 interacts with numerous proteins. Further studies on these interacting proteins may provide further insights into membrane dynamics during autophagy. Here, we identify Cis1/Atg31 as a protein that exhibits similar phenotypes to Atg17. ATG31 null cells were defective in autophagy and lost viability under starvation conditions. Localization of Atg31 to pre-autophagosomal structures (PAS) was dependent on Atg17. Coimmunoprecipitation experiments indicated that Atg31 interacts with Atg17. Together, Atg31 is a novel protein that, in concert with Atg17, is required for proper autophagosome formation.     

Atg15 in Saccharomyces cerevisiae consists of two functionally distinct domains

Autophagy is a cellular degradation system widely conserved among eukaryotes. During autophagy, cytoplasmic materials fated for degradation are compartmentalized in double membrane–bound organelles called autophagosomes. After fusing with the vacuole, their inner membrane–bound structures are released into the vacuolar lumen to become autophagic bodies and eventually degraded by vacuolar hydrolases. Atg15 is a lipase that is essential for disintegration of autophagic body membranes and has a transmembrane domain at the N-terminus and a lipase domain at the C-terminus. However, the roles of the two domains in vivo are not well understood. In this study, we found that the N-terminal domain alone can travel to the vacuole via the multivesicular body pathway, and that targeting of the C-terminal lipase domain to the vacuole is required for degradation of autophagic bodies. Moreover, we found that the C-terminal domain could disintegrate autophagic bodies when it was transported to the vacuole via the Pho8 pathway instead of the multivesicular body pathway. Finally, we identified H435 as one of the residues composing the putative catalytic triad and W466 as an important residue for degradation of autophagic bodies. This study may provide a clue to how the C-terminal lipase domain recognizes autophagic bodies to degrade them.     

Antibiotic Resistant Group A Streptococci

A series of Streptococcus pyogenes strains, including strains isolated from patients, mutants which had acquired in vitro resistance to penicillin (Pc), mitomycin C (MC), tetracycline (TC) and chloramphenicol (CM), ultraviolet light induced α hemolytic mutants, as well as β hemolytic mutants (β mutants) derived from α hemolytic mutants (α mutants) were compared as to their antibiotic sensitivity, and physiological, biochemical and serological properties. To obtain β mutants from α mutants the following procedures were employed: (1) serial mouse passage, (2) serial serum-broth transfers, (3) cultivation in heat-killed cultures of parent strains, and (4) cultivation in broth containing bacterial DNA extracted from parent streptococcus cells. From the results obtained these strains could be divided into two major groups, each with two subgroups. Group 1 strains produce soluble hemolysins and are sensitive to Pc. Subgroup 1–1 strains are sensitive to other antibiotics too; subgroup 1–2 are resistant to certain antibiotics other than Pc, bacitracin and MC. Group 2 strains do not produce soluble hemolysins and resistant to Pc. Subgroup 2-1 strains are α hemolytic on horse blood agar and subgroup 2–2 are β hemolytic on the same medium. Pc resistance in group 2 strains was more than 100-fold higher than that of sensitive strains, and was accompanied by MC resistance, but to a lesser degree. Pc resistance in group 2 mutants could be induced by antibiotics other than Pc and also by ultraviolet irradiation. Although group 1 cells retained the characteristics of typical S. pyogenes, group 2 cells, both α and β hemolytic, lost most of the physiological, biochemical and serological properties of this species. The similarity of group 2 strains to group D or group N streptococcal strains in their general properties is discussed.     

Transmission of Bacteriocinogenicity by Conjugation in Group D Streptococci

Seventy-seven out of eighty-one group D streptococcal strains isolated from humans and animals were found to produce bacteriocins that were active on other streptococcal strains of gorup A and D, but inactive on their own cells. On the bases of the spectra of indicator strains, and the sensitivities to heat, chloroform, and trypsin, seven types of bacteriocins were classified. Streptococcus faecalis var. liquefaciens strain 4532 or strain A (liq-A) was UV-irradiated, and mutants which lost bacteriocin- as well as the β-hemolysin-forming activities (Bact^–.Hem^–) were obtained. Cells of the type I bacteriocin producer (SM^r.TC^r.Bact-I⁺.Hem⁺) and nonproducer 2025 (PC^r.Bact-I^–.Hem^–), both belonging to S. faecalis var. liquefaciens, were mixed and incubated in broth. Recombinants (PC^r.-SM^s.TC^s.Bact-I⁺.Hem⁺) were obtained at a high frequency (5.8% preinoculum size of PC^r.-Bact-I^–.Hem^–), and the character was stable for at least ten transfers. In the mixed culture, a marked decrease in the recipient 2025 cell number was observed. The occurrence of recombinants was not inhibited by deoxyribonuclease. A cell-free filtrate of Bact⁺.Hem⁺ cells mixed with Bact^–.Hem^– cells did pot cause a mutation of the latter combined characters. The transfer of a genetic marker is discussed as an event of the cell-to-cell contact.     

Changes in Sulfhydryl Groups and Hydrodynamic Behavior of Thiolated Polyvinyl Alcohol Reacted with Oxidized Lipid

A synthetic polymer, thiolated polyvinyl alcohol (SH-PVA), was used for examining the reaction between sulfhydryls and lipids during dough mixing. When SH-PVA was incubated with linoleic acid hydroperoxide at a concentration of 1/12 to 1/4 mole to a mole of SH group, progressive oxidation of sulfhydryl was observed and a part of disulfides seemed to be further oxidized in some cases. The hydrodynamic behavior of SH-PVA treated with oxidized linoleic acid was similar to that of SH-PVA treated with an oxidizing agent, potassium bromate; non-Newtonian flow and increase in viscosity were observed. The peak of the gel filtration pattern of SH-PVA shifted to a faster elution position upon treatment with oxidized linoleic acid. These results probably indicate the formation of an interchain disulfide linkage. In view of these results, we discussed the role of lipids involved in oxidation of free SH groups in the rheological properties of gluten and dough.     

Transformability of Group H Streptococcus Challis

From a wild type strain Challis of the group H streptococcus, greening (Challis α) and β-hemolytic (Challis β) colonies were isolated on horse blood agar. Both colonies formed greening on sheep blood agar, and no significant differences were found in their biological, serological and chemical analyses. They, however, showed clear differences on the transformability. Transformability, the producibility of competence-provoking factor (CPF) and competency which have been reported on the Challis strain were all found in Challis β strain. On the other hand, Challis α strain did not produce antibiotic-resistant transformants with the addition of CPF, and could not produce CPF even when the cells were cultured under various conditions of incubation or treated with lysozyme or detergents. The transformabilities of antibiotic-resistant mutants obtained from the Challis β strain were lower than those of the original Challis β strain, as pointed out by other investigators, while the Challis α strain became transformable on antibiotic resistance only when it acquired streptomycin resistance. In the Challis β strain and the antibiotic-resistant mutants of Challis α strain, the separate markers of streptomycin, penicillin, tetracycline, mitomycin C, as well as the combinations of these markers were found to be transformed at the highest rate in the strains having transformation of streptomycin resistance. The findings are discussed with respect to incorporation of deoxyribonucleic acid into recipient cells and to the reports of other workers.     

Characteristics of Group A Streptococcal Variants Treated with Mitomycin C

Transfers of Streptococcus pyogenes strain T12 in Todd–Hewitt broth containing stepwise increases in amounts of mitomycin C (MC) gave rise to slight changes of their colonial appearances. Variants thus obtained were examined for antibiotic and bile resistances; production of streptolysin-S, -O and deoxyribonuclease; growth in alkaline medium, high salt concentration, and at 10 C and 45 C; sugar fermentations, and precipitin reactions. Four strains retained group A antigen, but some of them lost the ability to produce hemolysins and deoxyribonuclease, and acquired resistance to bile, penicillin and streptomycin as well as MC, and to physical environments. Four other strains lost group A antigen and acquired new antigens common to cells of group C, group D, or highly antibiotic-resistant mutants reported previously. A variant which reacted with group C antiserum contained galactosamine, but not glucosamine, while the parent strain showed the reverse pattern. Many other variants contained both hexosamines. Even a variant, strain TL3-2, reacted strongly only with group A antiserum, but contained glucose and both hexosamines. These strains having galactosamine possessed uridine diphosphate (UDP)-N-acetylglucosamine-4-epimerase activity which converted the substrate into UDP-N-acetylgalactosamine, while the parent strain failed to demonstrate the existence of this enzyme. The variants were discussed with respect to the group A streptococcal variations possessing no more original characteristics.