Programmable interactions and emergent geometry in an array of atom clouds.

Interactions govern the flow of information and the formation of correlations between constituents of many-body quantum systems, dictating phases of matter found in nature and forms of entanglement generated in the laboratory. Typical interactions decay with distance and thus produce a network of connectivity governed by geometry-such as the crystalline structure of a material or the trapping sites of atoms in a quantum simulator1,2. However, many envisioned applications in quantum simulation and computation require more complex coupling graphs including non-local interactions, which feature in models of information scrambling in black holes3-6 and mappings of hard optimization problems onto frustrated classical magnets7-11. Here we describe the realization of programmable non-local interactions in an array of atomic ensembles within an optical cavity, in which photons carry information between atomic spins12-19. By programming the distance dependence of the interactions, we access effective geometries for which the dimensionality, topology and metric are entirely distinct from the physical geometry of the array. As examples, we engineer an antiferromagnetic triangular ladder, a Möbius strip with sign-changing interactions and a treelike geometry inspired by concepts of quantum gravity5,20-22. The tree graph constitutes a toy model of holographic duality21,22, in which the quantum system lies on the boundary of a higher-dimensional geometry that emerges from measured correlations23. Our work provides broader prospects for simulating frustrated magnets and topological phases24, investigating quantum optimization paradigms10,11,25,26 and engineering entangled resource states for sensing and computation27,28.

Protecting Spin Coherence in a Tunable Heisenberg Model.

Using an ensemble of atoms in an optical cavity, we engineer a family of nonlocal Heisenberg Hamiltonians with continuously tunable anisotropy of the spin-spin couplings. We thus gain access to a rich phase diagram, including a paramagnetic-to-ferromagnetic Ising phase transition that manifests as a diverging magnetic susceptibility at the critical point. The susceptibility displays a symmetry between Ising interactions and XY (spin-exchange) interactions of the opposite sign, which is indicative of the spatially extended atomic system behaving as a single collective spin. Images of the magnetization dynamics show that spin-exchange interactions protect the coherence of the collective spin, even against inhomogeneous fields that completely dephase the noninteracting and Ising systems. Our results underscore prospects for harnessing spin-exchange interactions to enhance the robustness of spin squeezing protocols.

Three-Path Atom Interferometry with Large Momentum Separation.

We demonstrate the scale up of a symmetric three-path contrast interferometer to large momentum separation. The observed phase stability at separation of 112 photon recoil momenta exceeds the performance of earlier free-space interferometers. In addition to the symmetric interferometer geometry and Bose-Einstein condensate source, the robust scalability of our approach relies on the suppression of undesired diffraction phases through a careful choice of atom optics parameters. The interferometer phase evolution is quadratic with number of recoils, reaching a rate as high as 7×10^{7} rad/s. We discuss the applicability of our method towards a new measurement of the fine-structure constant and a test of QED.

Gyroscopic stabilization minimizes drag on Ruellia ciliatiflora seeds.

Fruits of Ruellia ciliatiflora (Acanthaceae) explosively launch small (2.5 mm diameter × 0.46 mm thick), disc-shaped seeds at velocities over 15 m s-1, reaching distances of up to 7 m. Through high-speed video analysis, we observe that seeds fly with extraordinary backspin of up to 1660 Hz. By modelling the seeds as spinning discs, we show that flying with backspin is stable against gyroscopic precession. This stable backspin orientation minimizes the frontal area during flight, decreasing drag force on the seeds and thus increasing dispersal distance. From high-speed video of the seeds' flight, we experimentally determine drag forces that are 40% less than those calculated for a sphere of the same volume and density. This reduces the energy costs for seed dispersal by up to a factor of five.

Total synthesis of a functional designer eukaryotic chromosome.

Rapid advances in DNA synthesis techniques have made it possible to engineer viruses, biochemical pathways and assemble bacterial genomes. Here, we report the synthesis of a functional 272,871-base pair designer eukaryotic chromosome, synIII, which is based on the 316,617-base pair native Saccharomyces cerevisiae chromosome III. Changes to synIII include TAG/TAA stop-codon replacements, deletion of subtelomeric regions, introns, transfer RNAs, transposons, and silent mating loci as well as insertion of loxPsym sites to enable genome scrambling. SynIII is functional in S. cerevisiae. Scrambling of the chromosome in a heterozygous diploid reveals a large increase in a-mater derivatives resulting from loss of the MATα allele on synIII. The complete design and synthesis of synIII establishes S. cerevisiae as the basis for designer eukaryotic genome biology.

Polyubiquitin-sensor proteins reveal localization and linkage-type dependence of cellular ubiquitin signaling.

Polyubiquitin chain topology is thought to direct modified substrates to specific fates, but this function-topology relationship is poorly understood, as are the dynamics and subcellular locations of specific polyubiquitin signals. Experimental access to these questions has been limited because linkage-specific inhibitors and in vivo sensors have been unavailable. Here we present a general strategy to track linkage-specific polyubiquitin signals in yeast and mammalian cells, and to probe their functions. We designed several high-affinity Lys63 polyubiquitin-binding proteins and demonstrate their specificity in vitro and in cells. We apply these tools as competitive inhibitors to dissect the polyubiquitin-linkage dependence of NF-κB activation in several cell types, inferring the essential role of Lys63 polyubiquitin for signaling via the IL-1ß and TNF-related weak inducer of apoptosis (TWEAK) but not TNF-α receptors. We anticipate live-cell imaging, proteomic and biochemical applications for these tools and extension of the design strategy to other polymeric ubiquitin-like protein modifications.

Assembling DNA fragments by USER fusion.

Recent advances in DNA synthesis technology make it possible to design and synthesize DNA fragments of several kb in size. However, the process of assembling the smaller DNA fragments into a larger DNA segment is still a cumbersome process. In this chapter, we describe the use of the uracil specific excision reaction (USER)-mediated approach for rapid and efficient assembly of multiple DNA fragments both in vitro and in vivo (using Escherichia coli). For USER fusion in vitro assembly, each of the individual building blocks (BBs), 0.75 kb in size (that are to be assembled), was amplified using the appropriate forward and reverse primers containing a single uracil (U) and DNA polymerase. The overlaps between adjoining BBs were 8-13 base pairs. An equimolar of the amplified BBs were mixed together and treated by USER enzymes to generate complementary 3' single-strand overhangs between adjoining BBs, which were then ligated and amplified simultaneously to generate the larger 3-kb segments. The assembled fragments were then cloned into plasmid vectors and sequenced to confirm their identity. For USER fusion in vivo assembly in E. coli, USER treatment of the BBs was performed in the presence of a synthetic plasmid, which had 8-13 base pair overlaps at the 5'-end of the 5' BB and at the 3'-end of the 3' BB in the mixture. The USER treated product was then transformed directly into E. coli to efficiently and correctly reconstitute the recombinant plasmid containing the desired target insert. The latter approach was also used to rapidly assemble three different target genes into a vector to form a new synthetic plasmid construct.

Assembling large DNA segments in yeast.

As described in a different chapter in this volume, the uracil-specific excision reaction (USER) fusion method can be used to assemble multiple small DNA fragments (∼0.75-kb size) into larger 3-kb DNA segments both in vitro and in vivo (in Escherichia coli). However, in order to assemble an entire synthetic yeast genome (Sc2.0 project), we need to be able to assemble these 3-kb pieces into larger DNA segments or chromosome-sized fragments. This assembly into larger DNA segments is carried out in vivo, using homologous recombination in yeast. We have successfully used this approach to assemble a 40-kb chromosome piece in the yeast Saccharomyces cerevisiae. A lithium acetate (LiOAc) protocol using equimolar amount of overlapping smaller fragments was employed to transform yeast. In this chapter, we describe the assembly of 3-kb fragments with an overlap of one building block (∼750 base pairs) into a 40-kb DNA piece.

The Build-a-Genome course.

Build-a-Genome is an intensive laboratory course at Johns Hopkins University that introduces undergraduates to the burgeoning field of synthetic biology. In addition to lectures that provide a comprehensive overview of the field, the course contains a unique laboratory component in which the students contribute to an actual, ongoing project to construct the first synthetic eukaryotic cell, a yeast cell composed of man-made parts. In doing so, the students acquire basic molecular biology skills and gain a truly "graduate student-like experience" in which they take ownership of their projects, troubleshoot their own experiments, present at frequent laboratory meetings, and are given 24-h access to the laboratory, albeit with all the guidance they will need to complete their projects during the semester. In this chapter, we describe the organization of the course and provide advice for anyone interested in starting a similar course at their own institution.

Specificity of the BRISC deubiquitinating enzyme is not due to selective binding to Lys63-linked polyubiquitin.

BRISC (Brcc36-containing isopeptidase complex) is a four-subunit deubiquitinating (DUB) enzyme that has a catalytic subunit, called Brcc36, that is a member of the JAMM/MPN(+) family of zinc metalloproteases. A notable feature of BRISC is its high specificity for cleaving Lys(63)-linked polyubiquitin. Here, we show that BRISC selectivity is not due to preferential binding to Lys(63)-linked polyubiquitin but is instead dictated by how the substrate isopeptide linkage is oriented within the enzyme active site. BRISC possesses a high affinity binding site for the ubiquitin hydrophobic surface patch that accounts for the bulk of the affinity between enzyme and substrate. Although BRISC can interact with either subunit of a diubiquitin conjugate, substrate cleavage occurs only when BRISC is bound to the hydrophobic patch of the distal (i.e. the "S1") ubiquitin at a ubiquitin-ubiquitin cleavage site. The importance of the Lys(63)-linked proximal (S1') ubiquitin was underscored by our finding that BRISC could not cleave the isopeptide bond joining a ubiquitin to a non-ubiquitin substrate. Finally, we also show that Abro1, another BRISC subunit, binds directly to Brcc36 and that the Brcc36-Abro1 heterodimer includes a minimal complex with Lys(63)-specific DUB activity.

Translational and reflectional priming invariance: a retrospective.

Biederman and Cooper (1991a) showed that the presentation of a briefly presented image of an object at one position in the visual field facilitated its identification, as assessed by naming speed and accuracy, several minutes later. The facilitation was unaffected by a translation or a reflection of the stimulus. A component of this priming was visual rather than basic-level conceptual or lexical in that there was less facilitation for an object with the same name (and basic-level class) but a different shape. The invariance of priming to view variables has stood up well over the years and appears to be a general phenomenon--as long as the original structural description can be readily resolved--in that it has also been observed for variations in size and orientation in depth. Although priming was unaffected by a change in position, we documented that there was explicit memory for the position (and orientation and size) of the stimulus. The existence of two forms of representation from the identical stimulus presentation--one invariant and the other dependent on view variables--poses a challenge as to what can be concluded about view invariance from single-unit activity.

K63-specific deubiquitination by two JAMM/MPN+ complexes: BRISC-associated Brcc36 and proteasomal Poh1.

An unusual deubiquitinating (DUB) activity exists in HeLa cell extracts that is highly specific for cleaving K63-linked but not K48-linked polyubiquitin chains. The activity is insensitive to both N-ethyl-maleimide and ubiquitin aldehyde, indicating that it lacks an active site cysteine residue, and gel filtration experiments show that it resides in a high molecular weight (approximately 600 kDa) complex. Using a biochemical approach, we found that the K63-specific DUB activity co-fractionated through seven chromatographic steps with three multisubunit complexes: the 19S (PA700) portion of the 26S proteasome, the COP9 signalosome (CSN) and a novel complex that includes the JAMM/MPN+ domain-containing protein Brcc36. When we analysed the individual complexes, we found that the activity was intrinsic to PA700 and the Brcc36 isopeptidase complex (BRISC), but that the CSN-associated activity was due entirely to an interaction with Brcc36. None of the complexes cleave K6, K11, K29, K48 or alpha-linked polyubiquitin, but they do cleave K63 linkages within mixed-linkage chains. Our results suggest that specificity for K63-linked polyubiquitin is a common property of the JAMM/MPN+ family of DUBs.

Evidence for bidentate substrate binding as the basis for the K48 linkage specificity of otubain 1.

Otubain 1 belongs to the ovarian tumor (OTU) domain class of cysteine protease deubiquitinating enzymes. We show here that human otubain 1 (hOtu1) is highly linkage-specific, cleaving Lys48 (K48)-linked polyubiquitin but not K63-, K29-, K6-, or K11-linked polyubiquitin, or linear alpha-linked polyubiquitin. Cleavage is not limited to either end of a polyubiquitin chain, and both free and substrate-linked polyubiquitin are disassembled. Intriguingly, cleavage of K48-diubiquitin by hOtu1 can be inhibited by diubiquitins of various linkage types, as well as by monoubiquitin. NMR studies and activity assays suggest that both the proximal and distal units of K48-diubiquitin bind to hOtu1. Reaction of Cys23 with ubiquitin-vinylsulfone identified a ubiquitin binding site that is distinct from the active site, which includes Cys91. Occupancy of the active site is needed to enable tight binding to the second site. We propose that distinct binding sites for the ubiquitins on either side of the scissile bond allow hOtu1 to discriminate among different isopeptide linkages in polyubiquitin substrates. Bidentate binding may be a general strategy used to achieve linkage-specific deubiquitination.

What types of visual recognition tasks are mediated by the neural subsystem that subserves face recognition?

Three divided visual field experiments tested current hypotheses about the types of visual shape representation tasks that recruit the cognitive and neural mechanisms underlying face recognition. Experiment 1 found a right hemisphere advantage for subordinate but not basic-level face recognition. Experiment 2 found a right hemisphere advantage for basic but not superordinate-level animal recognition. Experiment 3 found that inverting animals eliminates the right hemisphere advantage for basic-level animal recognition. This pattern of results suggests that the cognitive and neural mechanisms underlying face recognition are recruited when computational demands of a shape representation task are best served through the use of coordinate (rather than categorical) spatial relations.

Biochemical analysis of Angelman syndrome-associated mutations in the E3 ubiquitin ligase E6-associated protein.

Angelman syndrome is a severe neurological disorder characterized by mental retardation, absent speech, ataxia, seizures, and hyperactivity. The gene affected in this disorder is UBE3A, the gene encoding the E6-associated protein (E6AP) ubiquitin-protein ligase. Most patients have chromosomal deletions that remove the entire maternal allele of UBE3A. However, a small subset of patients have E6AP point mutations that result in single amino acid changes or short in-frame deletions that still allow translation of a full-length protein. By studying these point mutations in E6AP, we found a strong correlation between Angelman-associated mutations and a loss of E3 ubiquitin ligase activity. Interestingly the point mutations affect E6AP activity in different ways. Some mutant proteins cannot form thiol ester intermediates with ubiquitin, others retain the thiol ester formation activity but cannot efficiently transfer ubiquitin to a substrate, and still others are unstable in cells. Our results suggest that the loss of E6AP catalytic activity and likely the improper regulation of E6AP substrate(s) are important in the development of Angelman syndrome.

Qualitative differences in the representation of spatial relations for different object classes.

Two experiments investigated whether the representations used for animal, produce, and object recognition code spatial relations in a similar manner. Experiment 1 tested the effects of planar rotation on the recognition of animals and nonanimal objects. Response times for recognizing animals followed an inverted U-shaped function, whereas those for basic-level object recognition followed an M-shaped function (with a dip at 1800). Experiment 2 tested for laterality effects in the recognition of animals, produce, and objects. A right-hemisphere advantage was found for recognizing animals, whereas no hemispheric advantages were found for recognizing produce or objects. These results suggest that the recognition of animals with nonunique structural descriptions is mediated using coordinate spatial relations, whereas most forms of basic-level object recognition are mediated using categorical spatial relations.

Attentional coding of categorical relations in scene perception: evidence from the flicker paradigm.

The purpose of the present investigation was to determine whether the positions of objects in a scene are coded relative to one another categorically (i.e., above, below, or side of; Experiment 1) and to determine whether spatial position in scene perception is coded preattentively or only under focused attention (Experiment 2). In Experiment 1, participants viewed alternating versions of a scene in which one of the objects in the scene changed its categorical relationship to the closest object in the scene, changed only its metric relationship to the closest object in a scene, or appeared and disappeared. Participants were faster at detecting changes that disrupted categorical relations than at detecting changes that disrupted only metric relations. In Experiment 2, this categorical advantage still occurred even when participants were cued to the location of the change. These results suggest that categorical spatial relations are being coded in scene perception and that attention is required in order to encode spatial relations.