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Extremal properties of sequences of zeros of analytic functions in spaces with mixed norm

Functions whose sequences of absolute values of zeros have certain extremal properties are constructed in Bergmann spaces with powerlike weight and in more general spaces with mixed norm. These properties can be expressed in terms of subsequences with indices satisfying certain necessary and sufficient conditions. In particular, using these functions it is possible to distinguish between the zero sets of the spaces in question in some cases which have not previously been investigated. This reveals the significant role which subsequences of zeros in general play in the solution of this type of problems. Bibliography: 8 titles.

97 MATHEMATICS AND COMPUTING↗

Convergence of ray sequences of Frobenius-Padé approximants

Let σ-hat be a Cauchy transform of a possibly complex-valued Borel measure σ and {p_n} a system of orthonormal polynomials with respect to a measure μ, where supp(μ)∩supp(σ)=∅. An (m,n)th Frobenius-Padé approximant to σ-hat is a rational function P/Q, deg (P)≤m, deg (Q)≤n, such that the first m+n+1 Fourier coefficients of the remainder function Q σ-hat −P vanish when the form is developed into a series with respect to the polynomials p{sub n}. We investigate the convergence of the Frobenius-Padé approximants to σ-hat along ray sequences n/(n+m+1)→c>0, n−1≤m, when μ and σ are supported on intervals of the real line and their Radon-Nikodym derivatives with respect to the arcsine distribution of the corresponding interval are holomorphic functions. Bibliography: 30 titles. (paper)

97 MATHEMATICS AND COMPUTING↗

The Era GTPase recognizes the GAUCACCUCC sequence and binds helix 45 near the 3; end of 16S rRNA

Era, composed of a GTPase domain and a K homology domain, is essential for bacterial cell viability. It is required for the maturation of 16S rRNA and assembly of the 30S ribosomal subunit. We showed previously that the protein recognizes nine nucleotides (1531{sup AUCACCUCC}1539) near the 3{prime} end of 16S rRNA, and that this recognition stimulates GTP-hydrolyzing activity of Era. In all three kingdoms of life, the 1530{sup GAUCA}1534 sequence and helix 45 (h45) (nucleotides 1506-1529) are highly conserved. It has been shown that the 1530{sup GA}1531 to 1530{sup AG}1531 double mutation severely affects the viability of bacteria. However, whether Era interacts with G1530 and/or h45 and whether such interactions (if any) contribute to the stimulation of Era's GTPase activity were not known. Here, we report two RNA structures that contain nucleotides 1506-1542 (RNA301), one in complex with Era and GDPNP (GNP), a nonhydrolysable GTP-analogue, and the other in complex with Era, GNP, and the KsgA methyltransferase. The structures show that Era recognizes 10 nucleotides, including G1530, and that Era also binds h45. Moreover, GTPase assay experiments show that G1530 does not stimulate Era's GTPase activity. Rather, A1531 and A1534 are most important for stimulation and h45 further contributes to the stimulation. Although G1530 does not contribute to the intrinsic GTPase activity of Era, its interaction with Era is important for binding and is essential for the protein to function, leading to the discovery of a new cold-sensitive phenotype of Era.

59 BASIC BIOLOGICAL SCIENCES↗

Perplexing cooperative folding and stability of a low-sequence complexity, polyproline 2 protein lacking a hydrophobic core

The burial of hydrophobic side chains in a protein core generally is thought to be the major ingredient for stable, cooperative folding. Here, we show that, for the snow flea antifreeze protein (sfAFP), stability and cooperativity can occur without a hydrophobic core, and without α-helices or β-sheets. We report sfAFP has low sequence complexity with 46% glycine and an interior filled only with backbone H-bonds between six polyproline 2 (PP2) helices. However, the protein folds in a kinetically two-state manner and is moderately stable at room temperature. We believe that a major part of the stability arises from the unusual match between residue-level PP2 dihedral angle bias in the unfolded state and PP2 helical structure in the native state. Additional stabilizing factors that compensate for the dearth of hydrophobic burial include shorter and stronger H-bonds, and increased entropy in the folded state. These results extend our understanding of the origins of cooperativity and stability in protein folding, including the balance between solvent and polypeptide chain entropies.

37 INORGANIC, ORGANIC, PHYSICAL, AND ANALYTICAL CH↗

In silico evidence for sequence-dependent nucleosome sliding

Significance The dynamic compaction of DNA into chromatin is essential for gene expression. Errors during compaction are associated with numerous diseases. Several molecular factors are known to affect chromatin dynamics, but their relative importance and the interplay between them are poorly understood. A detailed molecular model is used here to examine chromatin dynamics at the level of its most fundamental building block, namely the nucleosome. Nucleosome dynamics are demonstrated to be encoded in the DNA sequence itself, and key fundamental factors are uncovered that can significantly alter these dynamics at the molecular level. The results serve to complete a hitherto unavailable description of nucleosome dynamics by introducing previously unappreciated molecular processes, with the potential to influence macroscopic chromatin structure and genetics.

59 BASIC BIOLOGICAL SCIENCES↗

Linking secondary metabolites to gene clusters through genome sequencing of six diverse Aspergillus species

Significance The genus of Aspergillus holds fungi relevant to plant and human pathology, food biotechnology, enzyme production, model organisms, and a selection of extremophiles. Here we present six whole-genome sequences that represent unexplored branches of the Aspergillus genus. The comparison of these genomes with previous genomes, coupled with extensive chemical analysis, has allowed us to identify genes for toxins, antibiotics, and anticancer compounds, as well as show that Aspergillus novofumigatus is potentially as pathogenic as Aspergillus fumigatus , and has an even more diverse set of secreted bioactive compounds. The findings are of interest to industrial biotechnology and basic research, as well as medical and clinical research.

59 BASIC BIOLOGICAL SCIENCES↗

Structure of the zinc-finger antiviral protein in complex with RNA reveals a mechanism for selective targeting of CG-rich viral sequences

Infection of animal cells by numerous viruses is detected and countered by a variety of means, including recognition of nonself nucleic acids. The zinc finger antiviral protein (ZAP) depletes cytoplasmic RNA that is recognized as foreign in mammalian cells by virtue of its elevated CG dinucleotide content compared with endogenous mRNAs. Here, we determined a crystal structure of a protein-RNA complex containing the N-terminal, 4-zinc finger human (h) ZAP RNA-binding domain (RBD) and a CG dinucleotide-containing RNA target. We report the structure reveals in molecular detail how hZAP is able to bind selectively to CG-rich RNA. Specifically, the 4 zinc fingers create a basic patch on the hZAP RBD surface. The highly basic second zinc finger contains a pocket that selectively accommodates CG dinucleotide bases. Structure guided mutagenesis, cross-linking immunoprecipitation sequencing assays, and RNA affinity assays show that the structurally defined CG-binding pocket is not required for RNA binding per se in human cells. However, the pocket is a crucial determinant of high-affinity, specific binding to CG dinucleotide-containing RNA. Moreover, variations in RNA-binding specificity among a panel of CG-binding pocket mutants quantitatively predict their selective antiviral activity against a CG-enriched HIV-1 strain. Overall, the hZAP RBD RNA structure provides an atomic-level explanation for how ZAP selectively targets foreign, CG-rich RNA.

59 BASIC BIOLOGICAL SCIENCES↗

Untangling the sequence of events during the S 2 → S 3 transition in photosystem II and implications for the water oxidation mechanism

In oxygenic photosynthesis, light-driven oxidation of water to molecular oxygen is carried out by the oxygen-evolving complex (OEC) in photosystem II (PS II). Recently, we reported the room-temperature structures of PS II in the four (semi)stable S-states, S 1 , S 2 , S 3 , and S 0 , showing that a water molecule is inserted during the S 2 → S 3 transition, as a new bridging O(H)-ligand between Mn1 and Ca. To understand the sequence of events leading to the formation of this last stable intermediate state before O 2 formation, we recorded diffraction and Mn X-ray emission spectroscopy (XES) data at several time points during the S 2 → S 3 transition. At the electron acceptor site, changes due to the two-electron redox chemistry at the quinones, Q A and Q B , are observed. At the donor site, tyrosine Y Z and His190 H-bonded to it move by 50 µs after the second flash, and Glu189 moves away from Ca. This is followed by Mn1 and Mn4 moving apart, and the insertion of O X (H) at the open coordination site of Mn1. This water, possibly a ligand of Ca, could be supplied via a “water wheel”-like arrangement of five waters next to the OEC that is connected by a large channel to the bulk solvent. XES spectra show that Mn oxidation (τ of ∼350 µs) during the S 2 → S 3 transition mirrors the appearance of O X electron density. This indicates that the oxidation state change and the insertion of water as a bridging atom between Mn1 and Ca are highly correlated.

59 BASIC BIOLOGICAL SCIENCES↗

DNA affinity purification sequencing and transcriptional profiling reveal new aspects of nitrogen regulation in a filamentous fungus

Significance Microorganisms have evolved transcriptional networks to prioritize utilization of available nutrient sources. For filamentous fungi, such as Neurospora crassa , this entails distinguishing between a variety of organic and inorganic nitrogen sources. Here, we transcriptionally profiled the response of N. crassa to a variety of nitrogen sources and used DNA affinity purification sequencing to characterize the role of regulatory genes and their direct downstream targets. We identified a transcription factor responsible for regulating genes involved in amino acid and mannose metabolism. By comparing the genes regulated by transcription factors that regulate specific nitrogen utilization pathways and transcription factors that regulate utilization of all nitrogen sources that require metabolic processing before utilization, we revealed aspects of the nitrogen regulatory network.

59 BASIC BIOLOGICAL SCIENCES↗

Deciphering the evolution of flavin-dependent monooxygenase stereoselectivity using ancestral sequence reconstruction

Controlling the selectivity of a reaction is critical for target-oriented synthesis. Accessing complementary selectivity profiles enables divergent synthetic strategies, but is challenging to achieve in biocatalytic reactions given enzymes’ innate preferences of a single selectivity. Thus, it is critical to understand the structural features that control selectivity in biocatalytic reactions to achieve tunable selectivity. Here, we investigate the structural features that control the stereoselectivity in an oxidative dearomatization reaction that is key to making azaphilone natural products. Crystal structures of enantiocomplementary biocatalysts guided the development of multiple hypotheses centered on the structural features that control the stereochemical outcome of the reaction; however, in many cases, direct substitutions of active site residues in natural proteins led to inactive enzymes. Ancestral sequence reconstruction (ASR) and resurrection were employed as an alternative strategy to probe the impact of each residue on the stereochemical outcome of the dearomatization reaction. These studies suggest that two mechanisms are active in controlling the stereochemical outcome of the oxidative dearomatization reaction: one involving multiple active site residues in AzaH and the other dominated by a single Phe to Tyr switch in TropB and AfoD. Moreover, this study suggests that the flavin-dependent monooxygenases (FDMOs) adopt simple and flexible strategies to control stereoselectivity, which has led to stereocomplementary azaphilone natural products produced by fungi. This paradigm of combining ASR and resurrection with mutational and computational studies showcases sets of tools for understanding enzyme mechanisms and provides a solid foundation for future protein engineering efforts.

Science & Technology - Other Topics↗

Dissecting neurofilament tail sequence-phosphorylation-structure relationships with multicomponent reconstituted protein brushes

Neurofilaments (NFs) are multisubunit, bottlebrush-shaped intermediate filaments abundant in the axonal cytoskeleton. Each NF subunit contains a long intrinsically disordered tail domain, which protrudes from the NF core to form a “brush” surrounding each NF. Precisely how the tails’ variable charge patterns and repetitive phosphorylation sites mediate their conformation within the brush remains an open question in axonal biology. We address this problem by grafting recombinant NF tail protein constructs NF-Light, -Medium, and -Heavy (NFL, NFM, and NFH) to surfaces, yielding protein brushes of defined stoichiometry that can be phosphorylated in vitro. Atomic force microscopy measurements reveal that brush height depends on composition monotonically but not always linearly for binary NFL:NFM or NFL:NFH systems, and that NFM-based brushes are highly extended, while brushes incorporating the much larger NFH are surprisingly compact even after multisite phosphorylation. Complementary self-consistent field theory (SCFT) predicts multilayer brush morphologies for NFM and phosphorylated NFH brushes. Further experiments and SCFT analysis with designed mutants reveal that N-terminal negative charges in the NFH tail repel phosphorylated residues to generate the multilayer morphology, while the C-terminal charge-neutral region contributes to multilayer brush morphology but not total brush height. Charge-shuffled NFM variants show that charge segregation promotes brush collapse near physiological ionic strengths. Collectively, this study supports a role for NFM in establishing a dynamic range for NF brush conformation, lending insight into previous in vitro and in vivo findings. More broadly, this work establishes a platform for dissecting contributions of disordered protein sequence to conformation at interfaces.

Science & Technology - Other Topics↗

The Replication Focus Targeting Sequence (RFTS) Domain Is a DNA-competitive Inhibitor of Dnmt1

Dnmt1 (DNA methyltransferase 1) is the principal enzyme responsible for maintenance of cytosine methylation at CpG dinucleotides in the mammalian genome. The N-terminal replication focus targeting sequence (RFTS) domain of Dnmt1 has been implicated in subcellular localization, protein association, and catalytic function. However, progress in understanding its function has been limited by the lack of assays for and a structure of this domain. Here, we show that the naked DNA- and polynucleosome-binding activities of Dnmt1 are inhibited by the RFTS domain, which functions by virtue of binding the catalytic domain to the exclusion of DNA. Kinetic analysis with a fluorogenic DNA substrate established the RFTS domain as a 600-fold inhibitor of Dnmt1 enzymatic activity. The crystal structure of the RFTS domain reveals a novel fold and supports a mechanism in which an RFTS-targeted Dnmt1-binding protein, such as Uhrf1, may activate Dnmt1 for DNA binding.

Syeda, Farisa↗

THE RED-SEQUENCE CLUSTER SURVEY-2 (RCS-2): SURVEY DETAILS AND PHOTOMETRIC CATALOG CONSTRUCTION

The second Red-sequence Cluster Survey (RCS-2) is a {approx}1000 deg{sup 2}, multi-color imaging survey using the square-degree imager, MegaCam, on the Canada-France-Hawaii Telescope. It is designed to detect clusters of galaxies over the redshift range 0.1 {approx}< z {approx}< 1. The primary aim is to build a statistically complete, large ({approx}10{sup 4}) sample of clusters, covering a sufficiently long redshift baseline to be able to place constraints on cosmological parameters via the evolution of the cluster mass function. Other main science goals include building a large sample of high surface brightness, strongly gravitationally lensed arcs associated with these clusters, and an unprecedented sample of several tens of thousands of galaxy clusters and groups, spanning a large range of halo mass, with which to study the properties and evolution of their member galaxies. This paper describes the design of the survey and the methodology for acquiring, reducing, and calibrating the data for the production of high-precision photometric catalogs. We describe the method for calibrating our griz imaging data using the colors of the stellar locus and overlapping Two Micron All Sky Survey photometry. This yields an absolute accuracy of <0.03 mag on any color and {approx}0.05 mag in the r-band magnitude, verified with respect to the Sloan Digital Sky Survey (SDSS). Our astrometric calibration is accurate to <<0.''3 from comparison with SDSS positions. RCS-2 reaches average 5{sigma} point-source limiting magnitudes of griz = [24.4, 24.3, 23.7, 22.8], approximately 1-2 mag deeper than the SDSS. Due to the queue-scheduled nature of the observations, the data are highly uniform and taken in excellent seeing, mostly FWHM {approx}< 0.''7 in the r band. In addition to the main science goals just described, these data form the basis for a number of other planned and ongoing projects (including the WiggleZ survey), making RCS-2 an important next-generation imaging survey.

79 ASTRONOMY AND ASTROPHYSICS↗

THE EVOLUTION OF DUSTY STAR FORMATION IN GALAXY CLUSTERS TO z = 1: SPITZER INFRARED OBSERVATIONS OF THE FIRST RED-SEQUENCE CLUSTER SURVEY

We present the results of an infrared (IR) study of high-redshift galaxy clusters with the MIPS camera on board the Spitzer Space Telescope. We have assembled a sample of 42 clusters from the Red-Sequence Cluster Survey-1 over the redshift range 0.3 < z < 1.0 and spanning an approximate range in mass of 10{sup 14-15} M {sub ☉}. We statistically measure the number of IR-luminous galaxies in clusters above a fixed inferred IR luminosity of 2 × 10{sup 11} M {sub ☉}, assuming a star forming galaxy template, per unit cluster mass and find it increases to higher redshift. Fitting a simple power-law we measure evolution of (1 + z){sup 5.1±1.9} over the range 0.3 < z < 1.0. These results are tied to the adoption of a single star forming galaxy template; the presence of active galactic nuclei, and an evolution in their relative contribution to the mid-IR galaxy emission, will alter the overall number counts per cluster and their rate of evolution. Under the star formation assumption we infer the approximate total star formation rate per unit cluster mass (ΣSFR/M {sub cluster}). The evolution is similar, with ΣSFR/M {sub cluster} ∼ (1 + z){sup 5.4±1.9}. We show that this can be accounted for by the evolution of the IR-bright field population over the same redshift range; that is, the evolution can be attributed entirely to the change in the in-falling field galaxy population. We show that the ΣSFR/M {sub cluster} (binned over all redshift) decreases with increasing cluster mass with a slope (ΣSFR/M{sub cluster}∼M{sub cluster}{sup -1.5±0.4}) consistent with the dependence of the stellar-to-total mass per unit cluster mass seen locally. The inferred star formation seen here could produce ∼5%-10% of the total stellar mass in massive clusters at z = 0, but we cannot constrain the descendant population, nor how rapidly the star-formation must shut-down once the galaxies have entered the cluster environment. Finally, we show a clear decrease in the number of IR-bright galaxies per unit optical galaxy in the cluster cores, confirming star formation continues to avoid the highest density regions of the universe at z ∼ 0.75 (the average redshift of the high-redshift clusters). While several previous studies appear to show enhanced star formation in high-redshift clusters relative to the field we note that these papers have not accounted for the overall increase in galaxy or dark matter density at the location of clusters. Once this is done, clusters at z ∼ 0.75 have the same or less star formation per unit mass or galaxy as the field.

79 ASTRONOMY AND ASTROPHYSICS↗

Orbital motion in pre-main sequence binaries

We present results from our ongoing program to map the visual orbits of pre-main sequence (PMS) binaries in the Taurus star forming region using adaptive optics imaging at the Keck Observatory. We combine our results with measurements reported in the literature to analyze the orbital motion for each binary. We present preliminary orbits for DF Tau, T Tau S, ZZ Tau, and the Pleiades binary HBC 351. Seven additional binaries show curvature in their relative motion. Currently, we can place lower limits on the orbital periods for these systems; full solutions will be possible with more orbital coverage. Five other binaries show motion that is indistinguishable from linear motion. We suspect that these systems are bound and might show curvature with additional measurements in the future. The observations reported herein lay critical groundwork toward the goal of measuring precise masses for low-mass PMS stars.

79 ASTRONOMY AND ASTROPHYSICS↗

THE EXTENDED MAIN-SEQUENCE TURNOFF CLUSTERS OF THE LARGE MAGELLANIC CLOUD-MISSING LINKS IN GLOBULAR CLUSTER EVOLUTION

Recent observations of intermediate-age (1-3 Gyr) massive star clusters in the Large Magellanic Cloud have revealed that the majority possess bifurcated or extended main-sequence turnoff (EMSTO) morphologies. This effect can be understood to arise from subsequent star formation among the stellar population with age differences between constituent stars amounting to 50-300 Myr. Age spreads of this order are similarly invoked to explain the light-element abundance variations witnessed in ancient globular clusters (GCs). In this paper, we explore the proposition that the clusters exhibiting the EMSTO phenomenon are a general phase in the evolution of massive clusters, one that naturally leads to the particular chemical properties of the ancient GC population. We show that the isolation of EMSTO clusters to intermediate ages is the consequence of observational selection effects. In our proposed scenario, the EMSTO phenomenon is identical to that which establishes the light-element abundance variations that are ubiquitous in the ancient GC population. Our scenario makes a strong prediction: EMSTO clusters will exhibit abundance variations in the light-elements characteristic of the ancient GC population.

79 ASTRONOMY AND ASTROPHYSICS↗

POPULATION PARAMETERS OF INTERMEDIATE-AGE STAR CLUSTERS IN THE LARGE MAGELLANIC CLOUD. III. DYNAMICAL EVIDENCE FOR A RANGE OF AGES BEING RESPONSIBLE FOR EXTENDED MAIN-SEQUENCE TURNOFFS

We present a new analysis of 11 intermediate-age (1-2 Gyr) star clusters in the Large Magellanic Cloud based on Hubble Space Telescope imaging data. Seven of the clusters feature main-sequence turnoff (MSTO) regions that are wider than can be accounted for by a simple stellar population, whereas their red giant branches (RGBs) indicate a single value of [Fe/H]. The star clusters cover a range in present-day mass from about 1 x 10{sup 4} M{sub sun} to 2 x 10{sup 5} M{sub sun}. We compare radial distributions of stars in the upper and lower parts of the MSTO region, and calculate cluster masses and escape velocities from the present time back to a cluster age of 10 Myr. Our main result is that for all clusters in our sample with estimated escape velocities v{sub esc} {approx}> 15 km s{sup -1} at an age of 10 Myr, the stars in the brightest half of the MSTO region are significantly more centrally concentrated than the stars in the faintest half and more massive RGB and asymptotic giant branch stars. This is not the case for clusters with v{sub esc} {approx}< 10 km s{sup -1} at an age of 10 Myr. We argue that the wide MSTO region of such clusters is caused mainly by a {approx}200-500 Myr range in the ages of cluster stars due to extended star formation within the cluster from material shed by first-generation stars featuring slow stellar winds. Dilution of this enriched material by accretion of ambient interstellar matter is deemed plausible if the spread of [Fe/H] in this ambient gas was very small when the second-generation stars were formed in the cluster.

79 ASTRONOMY AND ASTROPHYSICS↗

ON THE RELIABILITY OF STELLAR AGES AND AGE SPREADS INFERRED FROM PRE-MAIN-SEQUENCE EVOLUTIONARY MODELS

We revisit the problem of low-mass pre-main-sequence stellar evolution and its observational consequences for where stars fall on the Hertzsprung-Russell diagram (HRD). In contrast to most previous work, our models follow stars as they grow from small masses via accretion, and we perform a systematic study of how the stars' HRD evolution is influenced by their initial radius, by the radiative properties of the accretion flow, and by the accretion history, using both simple idealized accretion histories and histories taken from numerical simulations of star cluster formation. We compare our numerical results to both non-accreting isochrones and to the positions of observed stars in the HRD, with a goal of determining whether both the absolute ages and the age dispersions inferred from non-accreting isochrones are reliable. We show that non-accreting isochrones can sometimes overestimate stellar ages for more massive stars (those with effective temperatures above {approx}3500 K), thereby explaining why non-accreting isochrones often suggest a systematic age difference between more and less massive stars in the same cluster. However, we also find the only way to produce a similar overestimate for the ages of cooler stars is if these stars grow from {approx}0.01 M{sub sun} seed protostars that are an order of magnitude smaller than predicted by current theoretical models, and if the size of the seed protostar correlates systematically with the final stellar mass at the end of accretion. We therefore conclude that, unless both of these conditions are met, inferred ages and age spreads for cool stars are reliable, at least to the extent that the observed bolometric luminosities and temperatures are accurate. Finally, we note that the time dependence of the mass accretion rate has remarkably little effect on low-mass stars' evolution on the HRD, and that such time dependence may be neglected for all stars except those with effective temperatures above {approx}4000 K.

79 ASTRONOMY AND ASTROPHYSICS↗

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