Nucleotide sequences at the sites of action of the deoxyribonucleic acid modification enzyme of bacteriophage P1

Labelled oligonucleotides have been fractionated from DNAase digests of phage λ DNA that had been methylated with the phage P1 modification enzyme and S-[methyl-¹⁴C]adenosyl-l-methionine. The longest sequences established are the tetranucleotides pG-ǎ?-T-C4 and p?-T-C-T, which, together with the other sequences determined, particularly pA-G-?, show that the modification enzyme, M.EcoP1, methylates adenine residues within defined sequences and suggest that the oligonucleotide sequence recognized by this enzyme is the hexanucleotide pA-G-A-T-C-T. The duplex formed by base-pairing this hexanucleotide with its complementary sequence resembles the recognition sequence for several restriction enzymes in being bisected by an axis of 2-fold rotational symmetry.     

Synthesis of deoxyribonucleic acid in BHK-21/C13 cells

Newly synthesized (3)H-labelled DNA was extracted from baby hamster kidney cells (BHK-21/C13 cells) and was shown to possess single-stranded properties when examined by column chromatography on benzoylated naphthoylated DEAE-cellulose, hydroxyapatite and methylated albumin on kieselguhr, and by its affinity for nitrocellulose filters. Some of the newly synthesized DNA was shown to be of lower molecular weight than the bulk of the DNA when examined by alkaline sucrose-density-gradient centrifugation. The properties observed were not affected by treatment of the DNA with ribonuclease, Pronase or amylase. The effect of the size of the DNA on its observed properties was examined and is discussed. It is concluded that DNA synthesis in BHK-21/C13 cells proceeds according to the discontinuous-mechanism model in at least one of the strands.     

Using unmanned aerial vehicle (UAV) surveys and image analysis in the study of large surface‐associated marine species: a case study on reef sharks Carcharhinus melanopterus shoaling behaviour

A novel image analysis‐based technique applied to unmanned aerial vehicle (UAV) survey data is described to detect and locate individual free‐ranging sharks within aggregations. The method allows rapid collection of data and quantification of fine‐scale swimming and collective patterns of sharks. We demonstrate the usefulness of this technique in a small‐scale case study exploring the shoaling tendencies of blacktip reef sharks Carcharhinus melanopterus in a large lagoon within Moorea, French Polynesia. Using our approach, we found that C. melanopterus displayed increased alignment with shoal companions when distributed over a sandflat where they are regularly fed for ecotourism purposes as compared with when they shoaled in a deeper adjacent channel. Our case study highlights the potential of a relatively low‐cost method that combines UAV survey data and image analysis to detect differences in shoaling patterns of free‐ranging sharks in shallow habitats. This approach offers an alternative to current techniques commonly used in controlled settings that require time‐consuming post‐processing effort.     

Ephemeral ecological speciation and the latitudinal biodiversity gradient

The richness of biodiversity in the tropics compared to high‐latitude parts of the world forms one of the most globally conspicuous patterns in biology, and yet few hypotheses aim to explain this phenomenon in terms of explicit microevolutionary mechanisms of speciation and extinction. We link population genetic processes of selection and adaptation to speciation and extinction by way of their interaction with environmental factors to drive global scale macroecological patterns. High‐latitude regions are both cradle and grave with respect to species diversification. In particular, we point to a conceptual equivalence of “environmental harshness” and “hard selection” as eco‐evolutionary drivers of local adaptation and ecological speciation. By describing how ecological speciation likely occurs more readily at high latitudes, with such nascent species especially prone to extinction by fusion, we derive the ephemeral ecological speciation hypothesis as an integrative mechanistic explanation for latitudinal gradients in species turnover and the net accumulation of biodiversity.     

Assembly,molecular organization,and membrane-binding properties of development-specific septins

Septin complexes display remarkable plasticity in subunit composition, yet how a new subunit assembled into higher-order structures confers different functions is not fully understood. Here, this question is addressed in budding yeast, where during meiosis Spr3 and Spr28 replace the mitotic septin subunits Cdc12 and Cdc11 (and Shs1), respectively. In vitro, the sole stable complex that contains both meiosis-specific septins is a linear Spr28–Spr3–Cdc3–Cdc10–Cdc10–Cdc3–Spr3–Spr28 hetero-octamer. Only coexpressed Spr3 and Spr28 colocalize with Cdc3 and Cdc10 in mitotic cells, indicating that incorporation requires a Spr28-Spr3 protomer. Unlike their mitotic counterparts, Spr28-Spr3–capped rods are unable to form higher-order structures in solution but assemble to form long paired filaments on lipid monolayers containing phosphatidylinositol-4,5-bisphosphate, mimicking presence of this phosphoinositide in the prospore membrane. Spr28 and Spr3 fail to rescue the lethality of a cdc11Δ cdc12Δ mutant, and Cdc11 and Cdc12 fail to restore sporulation proficiency to spr3Δ/spr3Δ spr28Δ/spr28Δ diploids. Thus, specific meiotic and mitotic subunits endow septin complexes with functionally distinct properties.     

Marker Exchange Mutagenesis of mxaF,Encoding the Large Subunit of the Mxa Methanol Dehydrogenase,in Methylosinus trichosporium OB3b

Methanotrophs have remarkable redundancy in multiple steps of the central pathway of methane oxidation to carbon dioxide. For example, it has been known for over 30 years that two forms of methane monooxygenase, responsible for oxidizing methane to methanol, exist in methanotrophs, i.e., soluble methane monooxygenase (sMMO) and particulate methane monooxygenase (pMMO), and that expression of these two forms is controlled by the availability of copper. Specifically, sMMO expression occurs in the absence of copper, while pMMO expression increases with increasing copper concentrations. More recently, it was discovered that multiple forms of methanol dehydrogenase (MeDH), Mxa MeDH and Xox MeDH, also exist in methanotrophs and that the expression of these alternative forms is regulated by the availability of cerium. That is, expression of Xox MeDH increases in the presence of cerium, while Mxa MeDH expression decreases in the presence of cerium. As it had been earlier concluded that pMMO and Mxa MeDH form a supercomplex in which electrons from Mxa MeDH are back donated to pMMO to drive the initial oxidation of methane, we speculated that Mxa MeDH could be rendered inactive through marker-exchange mutagenesis but growth on methane could still be possible if cerium was added to increase the expression of Xox MeDH under sMMO-expressing conditions. Here we report that mxaF, encoding the large subunit of Mxa MeDH, could indeed be knocked out in Methylosinus trichosporium OB3b, yet growth on methane was still possible, so long as cerium was added. Interestingly, growth of this mutant occurred in both the presence and the absence of copper, suggesting that Xox MeDH can replace Mxa MeDH regardless of the form of MMO expressed.     

Differences in measures of boldness even when underlying behavioral syndromes are present in two populations of the song sparrow (<Emphasis Type="Italic">Melospiza melodia</Emphasis>)

One commonly studied behavioral syndrome is the correlation between aggression and boldness. Studies in song sparrows (M. melodia) have found greater aggression and boldness in urban populations and a correlation between aggression and boldness in rural populations, but not within urban populations. In previous studies, boldness was measured as flight initiation distance (FID), which may reflect habituation by urban birds to human presence. In this study, we measured boldness using playbacks of heterospecific alarm calls and investigated whether higher boldness is a general trait of urban birds and whether the same pattern of correlations between aggression and boldness would be seen. We conducted trials involving FID, alarm call playbacks and conspecific song playbacks on 25 birds from both an urban and a rural site. The results showed that urban birds were bolder, as measured by FID and response to alarm calls. Boldness and aggression were correlated in rural birds with each method of measuring boldness but were correlated in urban birds only when using alarm call playbacks. Our results suggest that a behavioral syndrome exists in both urban and rural populations but that urban birds are able to decrease their response to human disturbance.