Mapping and characterization of an E. coli mutant defective in IS1-mediated deletion formation

Summary A mutant of E. coli has been isolated in which the frequency of IS1-mediated deletion formation is reduced as much as 100 fold. The mutation causing this reduction, designated del, was mapped to a position near 61 min, in the vicinity of lysA and galR. Strains carrying a deletion of the del gene were constructed, and these exhibit a significant reduction in the frequency of IS1 excision in addition to impairment of deletion formation. A bacteriophage capable of transducing lysA and del was also isolated.     

A new host gene (groPC) necessary for lambda DNA replication.

Summary The isolation of a bacterial mutation in a gene, designated groPC, which affects the growth of phages lambda and P2 is described. Lambda replication is severely limited in the strain, and some lambda mutations, which map in (or near) the P gene, allow growth. The gro mutation, groPC259, is recessive to wild type and maps between threonine (thr) and diaminopimelate (dapB) on the E. coli chromosome. The possibility that the groPC gene is concerned with host DNA replication is discussed.     

A global synthesis of the rate and temperature sensitivity of soil nitrogen mineralization: latitudinal patterns and mechanisms

Soil net nitrogen (N) mineralization (N_min) is a pivotal process in the global N cycle regulating the N availability of plant growth. Understanding the spatial patterns of N_min, its temperature sensitivity (Q₁₀) and regulatory mechanisms is critical for improving the management of soil nutrients. In this study, we evaluated 379 peer‐reviewed scientific papers to explore how N_min and the Q₁₀ of N_min varied among different ecosystems and regions at the global scale. The results showed that N_min varied significantly among different ecosystems with a global average of 2.41 mg N soil kg^?1 day^?1. Furthermore, N_min significantly decreased with increasing latitude and altitude. The Q₁₀ varied significantly among different ecosystems with a global average of 2.21, ranging from the highest found in forest soils (2.43) and the lowest found for grassland soils (1.67) and significantly increased with increasing latitude. Path analyses indicated that N_min was primarily affected by the content of soil organic carbon (C), soil C:N ratio, and clay content, where Q₁₀ was primarily influenced by the soil C:N ratio and soil pH. Furthermore, the activation energy (E_a) of soil N mineralization was significantly and negative correlated with the substrate quality index among all ecosystems, indicating the applicability of the carbon quality temperature hypothesis to soil N mineralization at a global scale. These findings provided empirical evidence supporting that soil N availability, under global warming scenarios, is expected to increase stronger in colder regions as compared with that low‐latitude regions due to the higher Q₁₀. This may alleviate the restriction of N supply for increased primary productivity at higher latitudes.     

Suppression of polarity of insertion mutations within the gal operon of E. coli.

Summary Phenotypic revertants of galOP::IS1 and galOP::IS2 mutations have been isolated after mutagenesis with nitrosoguanidine, they are probably caused by mutations in gene suA. The polarity suppressor mutations described in this study and a known mutation in gene suA isolated by D. Morse (Morse and Guertin, 1972) suppress polarity caused by IS1 more effectively than that caused by IS2 or IS4. Furthermore, suppressibility is influenced by the site and orientation of IS integration.The synthesis of the three enzymes in galOP::IS suA double mutants is constitutive and the ratio of the three enzymes is altered in comparison to the wild type. The reasons for constitutive synthesis of the galactose enzymes and for the altered ratio of enzyme synthesis are discussed.     

Two kinds of insertions in bacterial genes

Summary Six insertion mutations in the gal operon of E. coli and two insertion mutations in the xycIIOP operon of bacteriophage lambda were tested for homology by annealing separated strands of lambda dgal DNA carrying the insertions, and inspection in the electron microscope.Class 1, consisting of the gal mutations OP 128, OP 141, T-N 116, OP 306, T-N 102 and the lambda mutation r14 are about 800 nucleotide pairs long, completely homologous and not circularly permuted. The first three insertions of class 1 are integrated in one direction with respect to the adjacent genes, the other three in the opposite direction. The DNA inserted in this class of mutations is called IS1.Class 2 consists of the gal insertion OP 308 and the lambda insertion r32. They are about 1400 nucleotide pairs long. The two are integrated in opposite direction with respect to the chromosome of dgal. The DNA in insertion mutations of class 2 will be called IS 2. IS1 and IS2 do not share any detectable homology.These data are supported by cross-hybridization experiments using RNA transcribed in vitro from lambda dgal or lambda DNA carrying one insertion and DNA carrying either the same or a different insertion.Similar results were obtained by Malamy, Fiandt, Szybalski and Fiandt, Szybalski, Malamy (accompanying papers).     

Linking elements to biochemicals: effects of nutrient supply ratios and growth rates on fatty acid composition of phytoplankton species

Three species of marine phytoplankton, Rhodomonas sp., Isochrysis galbana Parke, and Phaeodactylum tricornutum Bohlin, were cultivated in semicontinuous cultures to test biochemical responses (fatty acids; FAs) to five nitrogen (N):phosphorus (P) supply ratios and four growth rates (dilution rates). The characteristic FA profile was observed for each algal species (representing particular algal class), which remained relatively stable across the entire ranges of N:P supply ratios and growth rates. For all species, significant direct effects of N:P supply ratios on FAs were found at lower growth rates. The highest saturated and monounsaturated fatty acid (SFA and MUFA) contents were observed under N deficiency at the lowest growth rate in all three species, while responses of polyunsaturated fatty acids (PUFAs) revealed no consistent pattern. Total FAs (and SFAs and MUFAs) in all species showed significant negative correlations with N cell quota (Q_N) under N deficiency, but PUFAs had species‐specific correlations with Q_N. The results show that characteristic FA profiles of algal genus or species (representing particular algal classes) underlie fluctuations according to culture conditions. The significant correlation between FAs and Q_N under N deficiency suggests that elemental and biochemical limitation of phytoplankton should be considered mutually as determinants of food quality for zooplankton in marine ecosystems.     

The cis-specificity of the Q-gene product of bacteriophage lambda

Summary A rtrp/lac W205 substitution, fused to the late ragion of bacteriophage lambda, provided a convenient assay for phage late gene expression in the presence or absence of lambda pQ. Comparison of lacZ expression from Q ⁺ and Q ^- phages showed that late gene expression was markedly Q-dependent (263-fold difference). A cis/trans comparison of lambda pQ action showed a 180-fold difference in lacZ expression. The results suggest that pQ is only significantly active when supplied in cis to its site of action.     

DNA repair properties of Escherichia coli tif-1, recAo281 and lexA1 strains deficient in single-strand DNA binding protein

Summary Mutations affecting single-strand DNA binding protein (SSB) impair induction of mutagenic (SOS) repair. To further investigate the role of SSB in SOS induction and DNA repair, isogenic strains were constructed combining the ssb ⁺, ssb-1 or ssb-113 alleles with one or more mutations known to alter regulation of damage inducible functions. As is true in ssb ⁺ strains tif-1 (recA441) was found to allow thermal induction of prophage ⁺ and Weigle reactivation in ssb-1 and ssb-113 strains. Furthermore, tif-1 decreased the UV sensitivity of the ssb-113 strain slightly and permitted UV induction of prophage ⁺ at 30°C. Strains carrying the recAo281 allele were also constructed. This mutation causes high constitutive levels of RecA protein synthesis and relieves much of the UV sensitivity conferred by lexA ^– alleles without restoring SOS (error-prone) repair. In contrast, the recAo281 allele failed to alleviate the UV sensitivity associated with either ssb ^– mutation. In a lexA1 recAo281 background the ssb-1 mutation increased the extent of postirradiation DNA degradation and concommitantly increased UV sensitivity 20-fold to the level exhibited by a recA1 strain. The ssb-113 mutation also increased UV sensitivity markedly in this background but did so without greatly increasing postirradiation DNA degradation. These results suggest a direct role for SSB in recombinational repair apart from and in addition to its role in facilitating induction of the recA-lexA regulon.     

N-methyl-N′-nitro-N-nitrosoguanidine and UV induced mutants of the dinoflagellate Crypthecodinium cohnii*

SYNOPSIS. Mutant strains were chemically induced by treatment with N-methyl-N′-nitro-N-nitrosoguanidine (NTG) and UV irradiation. UV and NTG mutation rates were obtained that were both consistent with the organism being haploid. Three types of mutants were produced: (a) strains deficient in both β- and γ-carotene, the only carotenoids found in the wild type; phenotypes include albinos (translucent, dull white, “snow white”) and cream-colored on agar as compared to the yellow-orange color of wild type colonies; (b) strains requiring adenine, guanine or cytosine in addition to the minimal medium for growth; (c) mutants that grow at a rate less than 40% of the wild type in minimal medium.     

Spontaneous transposition in the bacteriophage lambda cro gene residing on a plasmid.

A new mutagenesis assay system based on the phage lambda cro repressor gene residing on a plasmid was developed. The assay detects mutations in cro that decrease the binding of the repressor to the OR operator in an OR PR-lacZ fusion present in a lambda prophage. Mutations arose spontaneously during growth of E. coli cells harboring cro plasmids at a frequency of 3-6 x 10(-6). Analysis of some 200 cro mutants from several 'wild-type' strains revealed a substantial fraction of 25-70% insertion events caused by transposition of IS elements. Most of the insertions were caused by IS1, but IS5 insertions were observed too. In strains harboring Tn10, IS10 was responsible for most insertions. Restriction nuclease digestion analysis revealed a preference for insertion of IS10 into the C-terminal half of cro, despite the absence of sequences which are known hot spots for Tn10 insertions. The frequency of IS1 insertions into cro decreased 25-60-fold and that of IS10 insertions decreased 200-fold in cells carrying the recA56 mutation, suggesting that RecA is involved in transposition of these elements. During the logarithmic phase of growth, the mutation frequency was constant for at least 22 generations; however, upon continuous incubation at the stationary phase, the mutation frequency gradually increased, yielding a 3-fold increase in the frequency of insertion and a 4-5-fold increase in point mutation. Genomic Southern analysis of chromosomal IS elements in cells which underwent a transposition from the chromosome into the cro plasmid revealed that the number and distribution of IS1 and IS5 were usually unaltered compared to cells which did not undergo a transposition event. In contrast, essentially each IS10 transposition was accompanied by multiple events which led to changes in the number and distribution of chromosomal IS10 elements.