Genetic and physical analyses of Caulobacter crescentus trp genes.

Caulobacter crescentus trp mutants were identified from a collection of auxotrophs. Precursor feeding experiments, accumulation studies, and complementation experiments resulted in the identification of six genes corresponding to trpA, trpB, trpC, trpD, trpE, and trpF. Genetic mapping experiments demonstrated that the trp genes were in two clusters, trpCDE and trpFBA, and a 5.4-kilobase restriction fragment from the C. crescentus chromosome was isolated that contained the trpFBA gene cluster. Complementation experiments with clones containing the 5.4-kilobase fragment indicated that trpF was expressed in Escherichia coli and that all three genes were expressed in Pseudomonas putida. This expression was lost in both organisms when the pBR322 tet gene promoter was inactivated, indicating that all three genes were transcribed in the same orientation from the tet promoter. Thus, the C. crescentus promoters do not seem to be expressed in E. coli or P. putida. Complementation of the C. crescentus trp mutants indicated that the tet promoter was not necessary for expression in C. crescentus and suggested that at least two native promoters were present for expression of the trpF, trpB, and trpA genes. Taken together, these results indicate that C. crescentus promoters may have structures that are significantly different from the promoters of other gram-negative species.     

Effects of nutritional environment and ram effect on breeding season in range sheep

Effects of season, nutritional environment, and presence of rams on incidence and rate of ovulation in fine-wool sheep in southern New Mexico were investigated over 14 months (February 1983-March 1984). Number of corpora lutea (CL) were observed each month by laparoscopy in random samples from six ewes in each subgroup. Seasonality of ovulation in fine-wool sheep managed on range was much more marked than in animals managed on alfalfa hay in drylot. Incidence of ovulation approached zero in range-managed ewes during May, June, and July (8%, 0%, 4%) but was higher (P<0.05) in the alfalfa-drylot group (42%, 17%, 75%), respectively. Although the incidence remained lower (P<0.05) in August in range-managed than in drylot ewes (50% and 83%, respectively), both groups were ovulating at a high rate during September through January. Incidence of ovulation dropped dramatically both years in the range-managed group in February compared with that of the alfalfa-drylot group (48% and 36% vs 100% and 83%). Mean ovulation rate per ewe ovulating did not differ (P>0.10) between nutrition groups within the 14 months. Body weight was related to CL number during periods of seasonal transition only. Continuous or intermittent presence of a ram had no real effect on either incidence or rate of ovulation during the transitional months. Nutritional environment can affect seasonality of breeding in fine-wool range sheep. This effect may not be consistently modified by presence of rams.     

The regulation of Mn2+ and Cu2+ uptake in cells of the yeast Candida utilis grown in continuous culture

Abstract Transport of Mn²⁺ was repressed in Candida utilis cells grown in continuous culture in high-Mn²⁺ (100 μM Mn²⁺) medium as compared to cells grown in basic (0.45 μM Mn²⁺) and low-Mn²⁺ (< 0.05 μM Mn²⁺) media. In contrast, no repression of Cu²⁺ uptake occurred in high-Cu²⁺-grown (25 μM Cu²⁺) cells as compared to cells grown in basic medium (0.54 μM Cu²⁺). Cu²⁺-limited cells did not hyperaccumulate Cu²⁺ and there was not significant difference in initial uptake rates for all 3 Cu²⁺ conditions. Mn²⁺ uptake appears to be regulated by a mechanism sensitive to the external Mn²⁺ concentration, whereas Cu²⁺ transport is not governed in this way by the external Cu²⁺.     

Selection in plant populations of effectively infinite size. V. Biallelic models of trioecy

A one-locus two-allele model of trioecy (presence of hermaphrodites, males and females in one population) is considered, in order to study the conditions for the persistence of this system. All possible assignments of the three sex types to the three genotypes are considered. This leads to three different modes of inheritance of trioecy, namely (a) females heterozygous, (b) males heterozygous and (c) hermaphrodites heterozygous, where in each mode each of the remaining two sex types is homozygous for one of the alleles. For mode (c) trioecy is always persistent, and the dependence of the sex ratio (for the three sex types) on the ovule and pollen fertilities and on the hermaphrodite selfing rate is specified. For the other two modes, (a) and (b), trioecy is not protected, i.e., it may not persist for any fertilities, viabilities or selfing rates. Thus, in this situation it is important to study the conditions under which the "marginal" systems of sexuality of trioecy, i.e., hermaphroditism, dioecy and gynodioecy in mode (a), and hermaphroditism, dioecy and androdioecy in mode (b), may become established. The results show that each marginal system may evolve from each other via trioecy. The evolution of dioecy is easier in mode (a) than in (b), so that female heterogamety would be expected to occur more often than male heterogamety in the present model. Under some conditions the breeding system obtained in equilibrium populations may depend on the initial genotype frequencies.—The necessity of considering modes of inheritance for sexual polymorphisms is demonstrated by comparing our results with those obtained from an evolutionary stable strategy (ESS) analysis of a purely phenotypic model.     

Arrangement of MP26 in lens junctional membranes: Analysis with proteases and antibodies

Summary The major membrane protein of the bovine lens fiber cell is a 26-kilodalton (kD) protein (MP26), which appears to be a component of the extensive junctional specializations found in these cells. To examine the arrangement of MP26 within the junctional membranes, various proteases were incubated with fiber cell membranes that had been isolated with or without urea and/or detergents. These membranes were analyzed with electron microscopy and SDS-PAGE to determine whether the junctional specializations or the proteins were altered by proteolysis. Microscopy revealed no obvious structural changes. Electrophoresis showed that chymotrypsin, papain, and trypsin degraded MP26 to 21–22 kD species. A variety of protease treatments, including overnight digestions, failed to generate additional proteolysis. Regions on MP26 which were sensitive to these three proteases overlapped. Smaller peptides were cleaved from MP26 with V8 protease and carboxypptidases A and B. Protein domains cleaved by these proteases also overlapped with regions sensitive to chymotrypsin, papain, and trypsin. Specific inhibition of the carboxypeptidases suggested that cleavage obtained with these preparations was not likely due to contaminating endoproteases. Since antibodies are not thought to readily penetrate the 2–3 nm extracellular gap in the fiber cell junctions, antibodies to MP26 were used to analyze the location of the protease-sensitive domains. Antisera were applied to control (26 kD) and proteolyzed (22 kD) membranes, with binding being evaluated by means of ELISA reactions on intact membranes. Antibody labeling was also done following SDS-PAGE and transfer to derivatized paper. Both assays showed a significant decrease in binding following proteolysis, with the 22 kD product showing no reaction with the anti-MP26 sera. These investigations suggest that MP26 is arranged with approximately fourfifths of the primary sequence “protected” by the lipid bilayer and the narrow extracellular gap. One-fifth of the molecule, including the C-terminus, appears to be exposed on the cytoplasmic side of the membrane.     

Selection in plant populations of effectively infinite size: IV. The maintenance of males among hermaphrodites for a biallelic model

Conditions for the maintenance of males in androdioecious populations (populations with both male and hermaphrodite individuals) have been derived for four different one-locus two-allele models of inheritance of androdioecy. The results are not in general accordance with those already known: depending on the mode of inheritance, males can be maintained irrespective of their fertilities. If males are sufficiently fertile, it may happen that they are maintained only for intermediate selfing rates of the hermaphrodites. A result already found for gynodioecy is confirmed for androdioecy, namely, that a 11 sex ratio is immediately established among zygotes if hermaphrodites appear as heterozygotes only.     

CELL CYCLE COMPARTMENT ANALYSIS OF CHINESE HAMSTER CELLS IN STATIONARY PHASE CULTURES

The distribution of Chinese hamster cells with respect to the compartments of the cell generation cycle was studied in cultures in the stationary phase of growth in two different media. A measure of the state of depletion of the nutrient medium was formulated by defining a quantity termed the nutritive capacity of the medium. This quantity was used to verify that the cessation of cell proliferation is due to nutrient deficiencies and not to density dependent growth inhibition. Cell cultures in stationary phase were diluted into fresh medium and as growth resumed, mitotic index, cumulative mitotic index, label index and viability were measured as a function of time. The distribution of cells with respect to compartments of the cell generation cycle in stationary phase populations was reconstructed from these data. Stationary phase populations of Chinese hamster cells that retained the capacity for renewed growth when diluted into fresh medium were found to be arrested in the G₁ and G₂ portions of the cycle; the relative proportion of these cells in G₁ increased with time in the stationary phase, but the sequence differs in the two media. In early stationary phase, in the less rich medium, more cells are in G₂ than in G₁. Also in this medium a fraction of the population was observed to be synthesizing DNA during stationary phase, but this fraction was not stimulated to renewed growth by dilution into fresh medium.