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Search indexed NASA NTRS and DOE OSTI research on propulsion, heat transfer, battery materials and energy systems. Follow report and document links to the original sources.

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At least 811 records · Page 45
All sourcesNASA NTRSDOE OSTI
DOE OSTI2011

Design of a shear-thinning recoverable peptide hydrogel from native sequences and application for influenza H1N1 vaccine adjuvant

Peptide hydrogels are considered injectable materials for drug delivery and tissue engineering applications. Most published hydrogel-forming sequences contain either alternating-charged and noncharged residues or amphiphilic blocks. Here, we report a self-assembling peptide, h9e (FLIVIGSIIGPGGDGPGGD), designed by rationally combining two native sequences from an elastic segment of spider silk and a trans-membrane segment of human muscle L-type calcium channel. The turning segment GSII of h9e promoted hydrogel formation in both Ca2+ solution and acidic pH conditions at water content greater than 99.5%. Although h9e Ca2+ hydrogel and h9e acidic hydrogel have the same sequence, they have distinct physical properties. The shear-thinning, rapid-strengthrecovering h9e Ca2+ hydrogel was used as an H1N1 influenza vaccine adjuvant. The h9e adjuvant was biologically safe and improved immune response by 70% compared with an oil-based commercial adjuvant.

59 BASIC BIOLOGICAL SCIENCES↗
DOE OSTI2018

Sequence isomeric giant surfactants with distinct self-assembly behaviors in solution

We have created and synthesized a pair of sequence isomeric giant surfactants based on polystyrene (PS) and polyhedral oligomeric silsesquioxane (POSS) nanoparticles. Despite the fact these two macromolecules possess identical compositions as “sequence isomers”, the distinctly arranged POSS sequences lead to different molecular packing conformations, and further induce distinguished self-assembly behaviors in DMF/water solutions.

37 INORGANIC, ORGANIC, PHYSICAL, AND ANALYTICAL CH↗
DOE OSTI2020

Decoding co-/post-transcriptional complexities of plant transcriptomes and epitranscriptome using next-generation sequencing technologies

Next-generation sequencing (NGS) technologies - Illumina RNA-seq, Pacific Biosciences isoform sequencing (PacBio Iso-seq), and Oxford Nanopore direct RNA sequencing (DRS) - have revealed the complexity of plant transcriptomes and their regulation at the co-/post-transcriptional level. Global analysis of mature mRNAs, transcripts from nuclear run-on assays, and nascent chromatin-bound mRNAs using short as well as full-length and single-molecule DRS reads have uncovered potential roles of different forms of RNA polymerase II during the transcription process, and the extent of co-transcriptional pre-mRNA splicing and polyadenylation. These tools have also allowed mapping of transcriptome-wide start sites in cap-containing RNAs, poly(A) site choice, poly(A) tail length, and RNA base modifications. The emerging theme from recent studies is that reprogramming of gene expression in response to developmental cues and stresses at the co-/post-transcriptional level likely plays a crucial role in eliciting appropriate responses for optimal growth and plant survival under adverse conditions. Although the mechanisms by which developmental cues and different stresses regulate co-/post-transcriptional splicing are largely unknown, a few recent studies indicate that the external cues target spliceosomal and splicing regulatory proteins to modulate alternative splicing. In this review, we provide an overview of recent discoveries on the dynamics and complexities of plant transcriptomes, mechanistic insights into splicing regulation, and discuss critical gaps in co-/post-transcriptional research that need to be addressed using diverse genomic and biochemical approaches.

Biochemistry & Molecular Biology↗
DOE OSTI2023

High-redshift quasars along the main sequence

The 4D Eigenvector 1 empirical formalism (4DE1) and its main sequence (MS) for quasars has emerged as a powerful tool for organising the diversity among quasar populations, as several key observational measures and physical parameters are systematically changing along it. Trends revealed by 4DE1 are very well established to explain all the diverse characteristics seen in low-redshift quasar samples. Nevertheless, the situation is far less clear when dealing with high-luminosity and high-redshift sources. Here, we aim to evaluate the behaviour of our sample of 22 quasars at high redshift (2.2 ≤ z ≤ 3.7) and high luminosity (47.39 ≤ L bol ≤ 48.36) in the context of the 4DE1. Our approach involves studying quasar physics through a spectroscopic exploration of UV and optical emission line diagnostics. We used new observations from the ISAAC instrument at ESO-VLT and primarily from the SDSS to cover the optical and the UV rest-frames, respectively. The emission lines were characterised both via a quantitative parametrisation of the line profiles and a decomposition of the emission line profiles using multicomponent fitting routines. We provide spectrophotometric properties and line profile measurements for Hβ+[O III]λλ4959,5007, as well as Si IVλ1397+O IV]λ1402, C IVλ1549+He IIλ1640, and the 1900 Å blend (including Al IIIλ1860, Si III]λ1892, and C III]λ1909). For six out of the 22 objects, a significantly blueshifted component on the Hβ profile is present. In 14 out of 22 cases, an Hβ outflowing component associated with [O III] is detected. The majority of [O III]λλ4959,5007 emission line profiles show blueshifted velocities higher than 250 km s -1 . We find extremely broad [O III]λλ4959,5007 emission that is comparable to the width of Hβ broad profile in some highly accreting quasars. The [O III]λλ4959,5007 and C IVλ1549 blueshifts show very high amplitudes and a high degree of correlation. The line widths and shifts are correlated for both [O III]λλ4959,5007 and C IVλ1549, suggesting that emission from outflowing gas is providing a substantial broadening effect to both lines. Otherwise, the links between C IVλ1549 centroid velocity at half intensity (c(1/2)), Eddington ratio (L/L Edd ), and bolometric luminosity are found to be in agreement with previous studies of high-luminosity quasars. Our analysis suggests that the behaviour of quasars of very high luminosity all along the main sequence is strongly affected by powerful outflows involving a broad range of spatial scales. The main sequence correlations remain valid at high redshift and high luminosity even if a systematic increase in line width is observed. Scaling laws based on UV Al IIIλ1860 and Hβ emission lines are equally reliable estimators of M BH .

79 ASTRONOMY AND ASTROPHYSICS↗
DOE OSTI2024

Towards a self-consistent model of the convective core boundary in upper main sequence stars: I. 2.5D and 3D simulations

There is strong observational evidence that the convective cores of intermediate-mass and massive main sequence stars are substantially larger than those predicted by standard stellar-evolution models. However, it is unclear what physical processes cause this phenomenon or how to predict the extent and stratification of stellar convective boundary layers. Convective penetration is a thermal-timescale process that is likely to be particularly relevant during the slow evolution on the main sequence. We use our low-Mach-number S EVEN -L EAGUE H YDRO code to study this process in 2.5D and 3D geometries. Starting with a chemically homogeneous model of a 15 M ⊙ zero-age main sequence star, we construct a series of simulations with the luminosity increased and opacity decreased by the same factor, ranging from 10 3 to 10 6 . After reaching thermal equilibrium, all of our models show a clear penetration layer; its thickness becomes statistically constant in time and it is shown to converge upon grid refinement. The penetration layer becomes nearly adiabatic with a steep transition to a radiative stratification in simulations at the lower end of our luminosity range. This structure corresponds to the adiabatic ‘step overshoot’ model often employed in stellar-evolution calculations. The simulations with the highest and lowest luminosity differ by less than a factor of two in the penetration distance. The high computational cost of 3D simulations makes our current 3D data set rather sparse. Depending on how we extrapolate the 3D data to the actual luminosity of the initial stellar model, we obtain penetration distances ranging from 0.09 to 0.44 pressure scale heights, which is broadly compatible with observations.

79 ASTRONOMY AND ASTROPHYSICS↗
DOE OSTI2011

Formation of laser-induced periodic surface structures on fused silica upon multiple cross-polarized double-femtosecond-laser-pulse irradiation sequences

The formation of laser-induced periodic surface structures (LIPSS) upon irradiation of fused silica with multiple irradiation sequences consisting of five Ti:sapphire femtosecond (fs) laser pulse pairs (150 fs, 800 nm) is studied experimentally. A Michelson interferometer is used to generate near-equal-energy double-pulse sequences with a temporal pulse delay from -20 to +20 ps between the cross-polarized individual fs-laser pulses ({approx}0.2 ps resolution). The results of multiple double-pulse irradiation sequences are characterized by means of Scanning Electron and Scanning Force Microscopy. Specifically in the sub-ps delay domain striking differences in the surface morphologies can be observed, indicating the importance of the laser-induced free-electron plasma in the conduction band of the solids for the formation of LIPSS.

36 MATERIALS SCIENCE↗
DOE OSTI2014

Sequence-dependent theory of oligonucleotide hybridization kinetics

A theoretical approach to the prediction of the sequence and temperature-dependent rate constants for oligonucleotide hybridization reactions has been developed based on the theory of relaxation kinetics. One-sided and two-sided melting reaction mechanisms for oligonucleotide hybridization reactions have been considered, analyzed, modified, and compared to select a physically consistent as well as robust model for prediction of the relaxation times of DNA hybridization reactions that agrees with the experimental evidence. The temperature- and sequence-dependent parameters of the proposed model have been estimated using available experimental data. The relaxation time model that we developed has been combined with the nearest neighbor model of hybridization thermodynamics to estimate the temperature- and sequence-dependent rate constants of an oligonucleotide hybridization reaction. The model-predicted rate constants are compared to experimentally determined rate constants for the same oligonucleotide hybridization reactions. Finally, we consider a few important applications of kinetically controlled DNA hybridization reactions.

37 INORGANIC, ORGANIC, PHYSICAL, AND ANALYTICAL CH↗
DOE OSTI2014

Dynamics and control of DNA sequence amplification

DNA amplification is the process of replication of a specified DNA sequence in vitro through time-dependent manipulation of its external environment. A theoretical framework for determination of the optimal dynamic operating conditions of DNA amplification reactions, for any specified amplification objective, is presented based on first-principles biophysical modeling and control theory. Amplification of DNA is formulated as a problem in control theory with optimal solutions that can differ considerably from strategies typically used in practice. Using the Polymerase Chain Reaction as an example, sequence-dependent biophysical models for DNA amplification are cast as control systems, wherein the dynamics of the reaction are controlled by a manipulated input variable. Using these control systems, we demonstrate that there exists an optimal temperature cycling strategy for geometric amplification of any DNA sequence and formulate optimal control problems that can be used to derive the optimal temperature profile. Strategies for the optimal synthesis of the DNA amplification control trajectory are proposed. Analogous methods can be used to formulate control problems for more advanced amplification objectives corresponding to the design of new types of DNA amplification reactions.

37 INORGANIC, ORGANIC, PHYSICAL, AND ANALYTICAL CH↗
DOE OSTI2015

Mesoscopic modeling of DNA denaturation rates: Sequence dependence and experimental comparison

Using rare event simulation techniques, we calculated DNA denaturation rate constants for a range of sequences and temperatures for the Peyrard-Bishop-Dauxois (PBD) model with two different parameter sets. We studied a larger variety of sequences compared to previous studies that only consider DNA homopolymers and DNA sequences containing an equal amount of weak AT- and strong GC-base pairs. Our results show that, contrary to previous findings, an even distribution of the strong GC-base pairs does not always result in the fastest possible denaturation. In addition, we applied an adaptation of the PBD model to study hairpin denaturation for which experimental data are available. This is the first quantitative study in which dynamical results from the mesoscopic PBD model have been compared with experiments. Our results show that present parameterized models, although giving good results regarding thermodynamic properties, overestimate denaturation rates by orders of magnitude. We believe that our dynamical approach is, therefore, an important tool for verifying DNA models and for developing next generation models that have higher predictive power than present ones.

37 INORGANIC, ORGANIC, PHYSICAL, AND ANALYTICAL CH↗
DOE OSTI2015

Anaerobic sequencing batch reactor in pilot scale for treatment of tofu industry wastewater

The small industry of tofu production process releases the waste water without being processed first, and the wastewater is directly discharged into water. In this study, Anaerobic Sequencing Batch Reactor in Pilot Scale for Treatment of Tofu Industry was developed through an anaerobic process to produce biogas as one kind of environmentally friendly renewable energy which can be developed into the countryside. The purpose of this study was to examine the fundamental characteristics of organic matter elimination of industrial wastewater with small tofu effective method and utilize anaerobic active sludge with Anaerobic Sequencing Bath Reactor (ASBR) to get rural biogas as an energy source. The first factor is the amount of the active sludge concentration which functions as the decomposers of organic matter and controlling selectivity allowance to degrade organic matter. The second factor is that HRT is the average period required substrate to react with the bacteria in the Anaerobic Sequencing Bath Reactor (ASBR).The results of processing the waste of tofu production industry using ASBR reactor with active sludge additions as starter generates cumulative volume of 5814.4 mL at HRT 5 days so that in this study it is obtained the conversion 0.16 L of CH{sub 4}/g COD and produce biogas containing of CH{sub 4}: 81.23% and CO{sub 2}: 16.12%. The wastewater treatment of tofu production using ASBR reactor is able to produce renewable energy that has economic value as well as environmentally friendly by nature.

09 BIOMASS FUELS↗
DOE OSTI2016

Time-optimal excitation of maximum quantum coherence: Physical limits and pulse sequences

Here we study the optimum efficiency of the excitation of maximum quantum (MaxQ) coherence using analytical and numerical methods based on optimal control theory. The theoretical limit of the achievable MaxQ amplitude and the minimum time to achieve this limit are explored for a set of model systems consisting of up to five coupled spins. In addition to arbitrary pulse shapes, two simple pulse sequence families of practical interest are considered in the optimizations. Compared to conventional approaches, substantial gains were found both in terms of the achieved MaxQ amplitude and in pulse sequence durations. For a model system, theoretically predicted gains of a factor of three compared to the conventional pulse sequence were experimentally demonstrated. Motivated by the numerical results, also two novel analytical transfer schemes were found: Compared to conventional approaches based on non-selective pulses and delays, double-quantum coherence in two-spin systems can be created twice as fast using isotropic mixing and hard spin-selective pulses. Also it is proved that in a chain of three weakly coupled spins with the same coupling constants, triple-quantum coherence can be created in a time-optimal fashion using so-called geodesic pulses.

71 CLASSICAL AND QUANTUM MECHANICS, GENERAL PHYSIC↗
DOE OSTI2017

Foldamer hypothesis for the growth and sequence differentiation of prebiotic polymers

It is not known how life originated. It is thought that prebiotic processes were able to synthesize short random polymers. However, then, how do short-chain molecules spontaneously grow longer? Also, how would random chains grow more informational and become autocatalytic (i.e., increasing their own concentrations)? We study the folding and binding of random sequences of hydrophobic ( H ) and polar ( P ) monomers in a computational model. We find that even short hydrophobic polar ( HP ) chains can collapse into relatively compact structures, exposing hydrophobic surfaces. In this way, they act as primitive versions of today’s protein catalysts, elongating other such HP polymers as ribosomes would now do. Such foldamer catalysts are shown to form an autocatalytic set, through which short chains grow into longer chains that have particular sequences. An attractive feature of this model is that it does not overconverge to a single solution; it gives ensembles that could further evolve under selection. This mechanism describes how specific sequences and conformations could contribute to the chemistry-to-biology (CTB) transition.

36 MATERIALS SCIENCE↗
DOE OSTI2011

IMPLICATIONS FOR THE BLAZAR SEQUENCE AND INVERSE COMPTON MODELS FROM FERMI BRIGHT BLAZARS

In this paper, we use the quasi-simultaneous spectra of Fermi bright blazars and Fermi-detected narrow-line Seyfert 1 (NLS1) to study the blazar sequence and inverse Compton models. (1) The synchrotron peak luminosities (L{sub s}) significantly correlate inversely with the synchrotron peak frequencies ({nu}{sub s}), L{sub s} {proportional_to}{nu}{sup -0.44{+-}0.11}{sub s}, which is consistent with the blazar sequence. In addition to the correlation, there are some blazars showing low {nu}{sub s} and low L{sub s} . To study the relation between these low-{nu}{sub s}-low-L{sub s} blazars and the blazar sequence, we present correlations of the parameter L{sub s} {nu}{sup 1/4}{sub s} with the ratio of Compton to synchrotron peak frequencies (r{sub Cs} {identical_to} {nu}{sub C}/{nu}{sub s}) and with the ratio of Compton to synchrotron luminosities (CD {identical_to} L{sub C} /L{sub s}). The results indicate that both correlations are significant with a Pearson's probability for a null correlation of p = 0.0218 and p = 0.0286, respectively. This does not support the idea that the low-{nu}{sub s}-low-L{sub s} blazars are sources with less beaming. Another possibility, as suggested by Ghisellini and Tavecchio, is that these blazars have relatively lower black hole masses. To test this, we collect the black hole masses of 30 blazars from archives and find that the black hole mass correlates with the parameter L{sub s} {nu}{sup 0.44}{sub s} (p = 0.0344). Therefore, the black hole masses of low-{nu}{sub s}-low-L{sub s} blazars are statistically small. The NLS1s are thought to have lower black hole masses. We find that the four NLS1s detected by Fermi have low {nu}{sub s} and low L{sub s} . This supports the above result. (2) The ratio r{sub Cs} correlates with CD significantly (p = 0.00375). The external Compton model can naturally explain this correlation, while the synchrotron self-Compton model cannot. This agrees with the findings of many authors that the EC process dominates the gamma-ray emission of flat spectrum radio quasars.

79 ASTRONOMY AND ASTROPHYSICS↗
DOE OSTI2012

EVOLUTION OF GROUP GALAXIES FROM THE FIRST RED-SEQUENCE CLUSTER SURVEY

We study the evolution of the red-galaxy fraction (f{sub red}) in 905 galaxy groups with 0.15 {<=} z < 0.52. The galaxy groups are identified by the 'probability friends-of-friends' algorithm from the first Red-Sequence Cluster Survey (RCS1) photometric-redshift sample. There is a high degree of uniformity in the properties of the red sequence of the group galaxies, indicating that the luminous red-sequence galaxies in the groups are already in place by z {approx} 0.5 and that they have a formation epoch of z {approx}> 2. In general, groups at lower redshifts exhibit larger f{sub red} than those at higher redshifts, showing a group Butcher-Oemler effect. We investigate the evolution of f{sub red} by examining its dependence on four parameters, one of which can be classified as intrinsic and three of which can be classified as environmental: galaxy stellar mass (M{sub *}), total group stellar mass (M{sub *,grp}, a proxy for group halo mass), normalized group-centric radius (r{sub grp}), and local galaxy density ({Sigma}{sub 5}). We find that M{sub *} is the dominant parameter such that there is a strong correlation between f{sub red} and galaxy stellar mass. Furthermore, the dependence of f{sub red} on the environmental parameters is also a strong function of M{sub *}. Massive galaxies (M{sub *} {approx}> 10{sup 11} M{sub Sun }) show little dependence of f{sub red} on r{sub grp}, M{sub *,grp}, and {Sigma}{sub 5} over the redshift range. The dependence of f{sub red} on these parameters is primarily seen for galaxies with lower masses, especially for M{sub *} {approx}< 10{sup 10.6} M{sub Sun }. We observe an apparent 'group down-sizing' effect, in that galaxies in lower-mass halos, after controlling for galaxy stellar mass, have lower f{sub red}. We find a dependence of f{sub red} on both r{sub grp} and {Sigma}{sub 5} after the other parameters are controlled. At a fixed r{sub grp}, there is a significant dependence of f{sub red} on {Sigma}{sub 5}, while r{sub grp} gradients of f{sub red} are seen for galaxies in similar {Sigma}{sub 5} regions. This indicates that galaxy group environment has a residual effect over that of local galaxy density (or vice versa), and both parameters need to be considered. This result suggests that processes identified with local galaxy density, such as galaxy harassment and mergers, and those associated with accretion into a larger group halo, such as ram pressure and strangulation, are both partaking in driving galaxies to their final red quiescent state. We discuss these results in the context of the 'nature versus nurture' scenario of galaxy evolution.

79 ASTRONOMY AND ASTROPHYSICS↗
DOE OSTI2013

Comparing the white dwarf cooling sequences in 47 Tuc and NGC 6397

Using deep Hubble Space Telescope imaging, color-magnitude diagrams are constructed for the globular clusters 47 Tuc and NGC 6397. As expected, because of its lower metal abundance, the main sequence of NGC 6397 lies well to the blue of that of 47 Tuc. A comparison of the white dwarf cooling sequences of the two clusters, however, demonstrates that these sequences are indistinguishable over most of their loci—a consequence of the settling out of heavy elements in the dense white dwarf atmosphere and the near equality of their masses. Lower quality data on M4 continues this trend to a third cluster whose metallicity is intermediate between these two. While the path of the white dwarfs in the color-magnitude diagram is nearly identical in 47 Tuc and NGC 6397, the numbers of white dwarfs along the path are not. This results from the relatively rapid relaxation in NGC 6397 compared to 47 Tuc and provides a cautionary note that simply counting objects in star clusters in random locations as a method of testing stellar evolutionary theory is likely dangerous unless dynamical considerations are included.

79 ASTRONOMY AND ASTROPHYSICS↗
DOE OSTI2013

Double blue straggler sequences in globular clusters: The case of NGC 362

We used high-quality images acquired with the Wide Field Camera 3 on board the Hubble Space Telescope to probe the blue straggler star (BSS) population of the galactic globular cluster NGC 362. We have found two distinct sequences of BSSs: this is the second case, after M30, where such a feature has been observed. Indeed, the BSS location, their extension in magnitude and color, and their radial distribution within the cluster nicely resemble those observed in M30, thus suggesting that the same interpretative scenario can be applied: the red BSS sub-population is generated by mass-transfer binaries, the blue one by collisions. The discovery of four new W UMa stars, three of which lie along the red BSS sequence, further supports this scenario. We also found that the inner portion of the density profile deviates from a King model and is well reproduced by either a mild power law (α ∼ –0.2) or a double King profile. This feature supports the hypothesis that the cluster is currently undergoing the core-collapse phase. Moreover, the BSS radial distribution shows a central peak and monotonically decreases outward without any evidence of an external rising branch. This evidence is a further indication of the advanced dynamical age of NGC 362; in fact, together with M30, NGC 362 belongs to the family of dynamically old clusters (Family III) in the 'dynamical clock' classification proposed by Ferraro et al. The observational evidence presented here strengthens the possible connection between the existence of a double BSS sequence and a quite advanced dynamical status of the parent cluster.

79 ASTRONOMY AND ASTROPHYSICS↗
DOE OSTI2011

SPITZER IRS SPECTRAL MAPPING OF THE TOOMRE SEQUENCE: SPATIAL VARIATIONS OF PAH, GAS, AND DUST PROPERTIES IN NEARBY MAJOR MERGERS

We have mapped the key mid-IR diagnostics in eight major merger systems of the Toomre sequence (NGC 4676, NGC 7592, NGC 6621, NGC 2623, NGC 6240, NGC 520, NGC 3921, and NGC 7252) using the Spitzer Infrared Spectrograph. With these maps, we explore the variation of the ionized-gas, polycyclic aromatic hydrocarbon (PAH), and warm gas (H{sub 2}) properties across the sequence and within the galaxies. While the global PAH interband strength and ionized gas flux ratios ([Ne III]/[Ne II]) are similar to those of normal star-forming galaxies, the distribution of the spatially resolved PAH and fine structure line flux ratios is significantly different from one system to the other. Rather than a constant H{sub 2}/PAH flux ratio, we find that the relation between the H{sub 2} and PAH fluxes is characterized by a power law with a roughly constant exponent (0.61 {+-} 0.05) over all merger components and spatial scales. While following the same power law on local scales, three galaxies have a factor of 10 larger integrated (i.e., global) H{sub 2}/PAH flux ratio than the rest of the sample, even larger than what it is in most nearby active galactic nuclei. These findings suggest a common dominant excitation mechanism for H{sub 2} emission over a large range of global H{sub 2}/PAH flux ratios in major mergers. Early-merger systems show a different distribution between the cold (CO J = 1-0) and warm (H{sub 2}) molecular gas components, which is likely due to the merger interaction. Strong evidence for buried star formation in the overlap region of the merging galaxies is found in two merger systems (NGC 6621 and NGC 7592) as seen in the PAH, [Ne II], [Ne III], and warm gas line emission, but with no apparent corresponding CO (J = 1-0) emission. The minimum of the 11.3/7.7 {mu}m PAH interband strength ratio is typically located in the nuclei of galaxies, while the [Ne III/[Ne II] ratio increases with distance from the nucleus. Our findings also demonstrate that the variations of the physical conditions within a merger are much larger than any systematic trends along the Toomre sequence.

79 ASTRONOMY AND ASTROPHYSICS↗
DOE OSTI2016

Coordinate action of distinct sequence elements localizes checkpoint kinase Hsl1 to the septin collar at the bud neck in Saccharomyces cerevisiae

Passage through the eukaryotic cell cycle requires processes that are tightly regulated both spatially and temporally. Surveillance mechanisms (checkpoints) exert quality control and impose order on the timing and organization of downstream events by impeding cell cycle progression until the necessary components are available and undamaged and have acted in the proper sequence. In budding yeast, a checkpoint exists that does not allow timely execution of the G2/M transition unless and until a collar of septin filaments has properly assembled at the bud neck, which is the site where subsequent cytokinesis will occur. An essential component of this checkpoint is the large (1518-residue) protein kinase Hsl1, which localizes to the bud neck only if the septin collar has been correctly formed. Hsl1 reportedly interacts with particular septins; however, the precise molecular determinants in Hsl1 responsible for its recruitment to this cellular location during G2 have not been elucidated. We performed a comprehensive mutational dissection and accompanying image analysis to identify the sequence elements within Hsl1 responsible for its localization to the septins at the bud neck. Unexpectedly, we found that this targeting is multipartite. A segment of the central region of Hsl1 (residues 611–950), composed of two tandem, semiredundant but distinct septin-associating elements, is necessary and sufficient for binding to septin filaments both in vitro and in vivo. However, in addition to 611–950, efficient localization of Hsl1 to the septin collar in the cell obligatorily requires generalized targeting to the cytosolic face of the plasma membrane, a function normally provided by the C-terminal phosphatidylserine-binding KA1 domain (residues 1379–1518) in Hsl1 but that can be replaced by other, heterologous phosphatidylserine-binding sequences.

59 BASIC BIOLOGICAL SCIENCES↗