Magnification of Genes Coding for Ribosomal RNA in Saccharomyces cerevisiae

When a strain of Saccharomyces cerevisiae monosomic for chromosome I and initially deficient for 25% of the genes coding for ribosomal RNA is repeatedly subcultured, the number of these genes increases to and remains stable at the number in the wild type. This strain shows 2:2; viable: inviable first division segregation and hemizygosity for the ade1 gene (a chromosome I marker), evidence that the strain is still monosomic for chromosome I. The increase in the number of genes coding for ribosomal RNA in yeast may be analogous to the magnification of the ribosomal RNA genes in Drosophila melanogaster bobbed mutants

Lessons from Ellen: A Case Study Investigation of Comprehension Strategy Instruction in Action

In the past ten years, reading comprehension instruction has received significant attention from educational researchers. Drawing on studies from cognitive psychology, reader response theory, and language arts research, current best practice in reading comprehension instruction is characterized by a strategies approach in which students are taught to think like proficient readers who visualize, infer, activate schema, question, and summarize as they read. Studies investigating the impact of comprehension strategy instruction on student achievement in reading suggest that when implemented consistently the intervention has a positive effect on achievement. Research also shows, however, that few teachers embrace this approach to reading instruction despite its effectiveness, even when the conditions for substantive professional development (i.e. prolonged engagement, support, resources, time) are present. The interpretive case study reported in this dissertation examined the year-long experience of one fourth grade teacher, Ellen, as she leanled about comprehension strategy instruction and attempted to integrate the approach in her reading program. The goal of the study was to extend current understanding of the factors that support or inhibit an individual teacher\u27s instructional decision making. The research explored how Ellen\u27s academic preparation, beliefs about reading comprehension instruction, and attitudes toward teacher-student interaction influenced her efforts to employ comprehension strategy instruction. Qualitative methods were the basis of this study\u27s research design. The primary methods for collecting data included pre- and post-interviews, field notes from classroom observations and staff development sessions, infonnal interviews, e-mail correspondence, and artifacts such as reading assignments, professional writing, school newsletters, and photographs of the classroom. Transcripts from interviews, as well as field notes, e-mail, and artifacts, were analyzed according to grounded theory\u27s constant-comparative method. The results of the study suggest that three factors were pivotal in Ellen\u27s successful implementation of reading strategy instruction: Pedagogical beliefs, classroom relationships, and professional community. Research on instructional change generally focuses on issues of time, resources, feedback, and follow-through. The research reported here recognizes the importance of these components, but expands contemporary thinking by showing how, in Ellen\u27s case, a teacher\u27s existing theories, her relationship with her students, and her professional interaction with peers impact instructional decisions

The proton electrochemical gradient induces a kinetic asymmetry in the symport cycle of LacY.

LacY catalyzes accumulation of galactosides against a concentration gradient by coupling galactoside and H+ transport (i.e., symport). While alternating access of sugar- and H+-binding sites to either side of the membrane is driven by binding and dissociation of sugar, the electrochemical H+ gradient ([Formula: see text]) functions kinetically by decreasing the Km for influx 50- to 100-fold with no change in Kd The affinity of protonated LacY for sugar has an apparent pK (pKapp) of ∼10.5, due specifically to the pKa of Glu325, a residue that plays an irreplaceable role in coupling. In this study, rates of lactose/H+ efflux were measured from pH 5.0 to 9.0 in the absence or presence of a membrane potential (ΔΨ, interior positive), and the effect of the imposed ΔΨ on the kinetics of efflux was also studied in right-side-out membrane vesicles. The findings reveal that [Formula: see text] induces an asymmetry in the transport cycle based on the following observations: 1) the efflux rate of WT LacY exhibits a pKapp of ∼7.2 that is unaffected by the imposed ΔΨ; 2) ΔΨ increases the rate of efflux at all tested pH values, but enhancement is almost 2 orders of magnitude less than observed for influx; 3) mutant Glu325 - Ala does little or no efflux in the absence or presence of ΔΨ, and ambient pH has no effect; and 4) the effect of ΔΨ (interior positive) on the Km for efflux is almost insignificant relative to the 50- to 100-fold decrease in the Km for influx driven by ΔΨ (interior negative)

Role of YidC in folding of polytopic membrane proteins

YidC of Echerichia coli, a member of the conserved Alb3/Oxa1/YidC family, is postulated to be important for biogenesis of membrane proteins. Here, we use as a model the lactose permease (LacY), a membrane transport protein with a known three-dimensional structure, to determine whether YidC plays a role in polytopic membrane protein insertion and/or folding. Experiments in vivo and with an in vitro transcription/translation/insertion system demonstrate that YidC is not necessary for insertion per se, but plays an important role in folding of LacY. By using the in vitro system and two monoclonal antibodies directed against conformational epitopes, LacY is shown to bind the antibodies poorly in YidC-depleted membranes. Moreover, LacY also folds improperly in proteoliposomes prepared without YidC. However, when the proteoliposomes are supplemented with purified YidC, LacY folds correctly. The results indicate that YidC plays a primary role in folding of LacY into its final tertiary conformation via an interaction that likely occurs transiently during insertion into the lipid phase of the membrane