Targeted Insertion of the mPing Transposable Element

Class II DNA Transposable Elements (TEs) are moved from one location to another in the genome by the action of transposase proteins that bind to repeat sequences at the ends of the elements. Although the location TE insertion is mostly random, the addition of DNA binding domains to the transposase proteins has allowed for targeted insertion of some elements. In this study, the Gal4 binding domain was added to the transposase proteins, ORF1 and TPase, which mobilize the mPing element from rice. The Gal4:TPase construct was capable of increasing the number of mPing insertions into the Gal2 and Gal4 promoter sequences in yeast. While this confirms that mPing insertion preference can be manipulated, the target specificity is relatively low. Thus, the CRISPR/Cas9 system was tested for its ability to generate targeted insertion of mPing. A dCas9:TPase fusion protein had a low transposition rate suggesting that the addition of this large protein disrupts TPase function. Unfortunately, the use of a MS2 binding domain to localize the TPase to the MS2 hairpin containing gRNA failed to produce targeted insertion. Thus, our results suggest that the addition of small DNA binding domain to the N-terminal of TPase is the best strategy for targeted insertion of mPing

The role of leadership in emergent, self-organization

As complex systems, organizations exist far from equilibrium where the ongoing interaction of system components leads to emergent and self-organizing behavior. What, then, is the role of leadership in systems where change often emerges in unexpected ways? In this paper, we build on the work of Marion and Uhl-Bien who suggest that in complex systems leaders enable rather than control the future. While traditional views of leadership focus on the leader’s responsibility for determining and directing the future through heavy reliance on control mechanisms, we offer empirical support for a different view of leadership based on a complexity perspective of organizations. Our findings show that as enablers, leaders disrupt existing patterns of behavior, encourage novelty, and make sense of emerging events for others. The results of our qualitative study include a set of research propositions as well as a discussion of the implications for managers and researchers

Structure and Mechanism of a Metal-Sensing Regulatory RNA

SummaryOrganisms maintain the correct balance of intracellular metals primarily through metal-sensing proteins that control transport and storage of the target ion(s). Here, we reveal the basis of metal sensing and genetic control by a metalloregulatory RNA. Our data demonstrate that a previously uncharacterized orphan riboswitch, renamed the “M-box,” is a divalent metal-sensing RNA involved in Mg2+ homeostasis. A combination of genetic, biochemical, and biophysical techniques demonstrate that Mg2+ induces a compacted tertiary architecture for M-box RNAs that regulates the accessibility of nucleotides involved in genetic control. Molecular details are provided by crystallographic structure determination of a Mg2+-bound M-box RNA. Given the distribution of this RNA element, it may constitute a common mode for bacterial metal ion regulation, and its discovery suggests the possibility of additional RNA-based metal sensors in modern and primordial organisms

Radical Change Accidentally: The Emergence and Amplification of Small Change

A decision to offer breakfast to homeless people led to radical change in a church and its environment. Existing theories of change do not fully explain observations from our qualitative study; however, complexity theory constructs suggest how and why such change emerged. We offer four key findings. First, the radical change was unintended, emergent, and slow. Second, destabilizing conditions helped small changes to emerge and become radical. Third, subsequent actions amplified an initial small change and, though not intended to do so, promoted radical change. Finally, the dynamic interaction of amplifying actions, contextual conditions, and small changes led to continuous radical change

Radical Change Accidentally: The Emergence and Amplification of Small Change

A decision to offer breakfast to homeless people led to radical change in a church and its environment. Existing theories of change do not fully explain observations from our qualitative study; however, complexity theory constructs suggest how and why such change emerged. We offer four key findings. First, the radical change was unintended, emergent, and slow. Second, destabilizing conditions helped small changes to emerge and become radical. Third, subsequent actions amplified an initial small change and, though not intended to do so, promoted radical change. Finally, the dynamic interaction of amplifying actions, contextual conditions, and small changes led to continuous radical change

De novo design of bioactive protein switches.

Allosteric regulation of protein function is widespread in biology, but is challenging for de novo protein design as it requires the explicit design of multiple states with comparable free energies. Here we explore the possibility of designing switchable protein systems de novo, through the modulation of competing inter- and intramolecular interactions. We design a static, five-helix 'cage' with a single interface that can interact either intramolecularly with a terminal 'latch' helix or intermolecularly with a peptide 'key'. Encoded on the latch are functional motifs for binding, degradation or nuclear export that function only when the key displaces the latch from the cage. We describe orthogonal cage-key systems that function in vitro, in yeast and in mammalian cells with up to 40-fold activation of function by key. The ability to design switchable protein functions that are controlled by induced conformational change is a milestone for de novo protein design, and opens up new avenues for synthetic biology and cell engineering

Renewed investigations at Taung; 90 years after the discovery of Australopithecus africanus

2015 marked the 90th anniversary of the description of the first fossil ofAustralopithecus africanus, commonly known as the Taung Child, which was unearthed during blasting at the Buxton-Norlim Limeworks (referred to as the BNL) 15 km SE of the town of Taung, South Africa. Subsequently, this site has been recognized as a UNESCOWorld Heritage site on the basis of its importance to southern African palaeoanthropology. Some other sites such as Equus Cave and Black Earth Cave have also been investigated; but the latter not since the 1940s. These sites indicate that the complex of palaeontological and archaeological localities at the BNL preserve a time sequence spanning the Pliocene to the Holocene. The relationship of these various sites and how they fit into the sequence of formation of tufa, landscapes and caves at the limeworks have also not been investigated or discussed in detail since Peabody’s efforts in the 1940s. In this contribution we mark the 90th anniversary of the discovery and description of the Taung Child by providing a critical review of previous work at Taung based on our recent preliminary work at the site. This includes a reassessment of the Taung Child Type Site, as well as renewed excavations at Equus Cave and the lesser-known locality and little-investigated Black Earth Cave. Preliminary results suggest that much of our previous understandings of the BNL’s formational history and site formation processes need to be reassessed. Only through detailed analysis on the BNLas a whole can we understand this complex depositional environment.Australian Research Council Future Fellowship grant FT120100399 Palaeontological Scientific Trust (PAST) National Geographic grants (8774-10 and 3212)JNC2016https://www.wits.ac.za/esi/palaeontologia-africana

Learning deficit in cognitively normal apoe ε4 carriers with low β-amyloid

Introduction: In cognitively normal (CN) adults, increased rates of amyloid beta (Aβ) accumulation can be detected in low Aβ (Aβ–) apolipoprotein E (APOE) ε4 carriers. We aimed to determine the effect of ε4 on the ability to benefit from experience (ie, learn) in Aβ–CNs. Methods: Aβ– CNs(n= 333) underwent episodic memory assessments every 18 months for 108 months. A subset (n = 48) completed the Online Repeatable Cognitive Assessment-Language Learning Test (ORCA-LLT) over 6 days. Results: Aβ– ε4 carriers showed significantly lower rates of improvement on episodic memory over 108 months compared to non-carriers (d = 0.3). Rates of learning on the ORCA-LLT were significantly slower in Aβ– ε4 carriers compared to non-carriers (d = 1.2). Discussion: In Aβ– CNs,ε4 is associated with a reduced ability to benefit from experience. This manifested as reduced practice effects (small to moderate in magnitude) over 108 months on the episodic memory composite, and a learning deficit (large in magnitude) over 6 days on the ORCA-LLT. Alzheimer’s disease (AD)–related cognitive abnormalities can manifest before preclinical AD thresholds

Drimolen cranium DNH 155 documents microevolution in an early hominin species

Paranthropus robustus is a small-brained extinct hominin from South Africa characterized by derived, robust craniodental morphology. The most complete known skull of this species is DNH 7 from Drimolen Main Quarry, which differs from P. robustus specimens recovered elsewhere in ways attributed to sexual dimorphism. Here, we describe a new fossil specimen from Drimolen Main Quarry, dated from approximately 2.04–1.95 million years ago, that challenges this view. DNH 155 is a well-preserved adult male cranium that shares with DNH 7 a suite of primitive and derived features unlike those seen in adult P. robustus specimens from other chronologically younger deposits. This refutes existing hypotheses linking sexual dimorphism, ontogeny and social behaviour within this taxon, and clarifies hypotheses concerning hominin phylogeny. We document small-scale morphological changes in P. robustus associated with ecological change within a short time frame and restricted geography. This represents the most highly resolved evidence yet of microevolutionary change within an early hominin species

Mutational analyses of UPIIIA, SHH, EFNB2, and HNF1β in persistent cloaca and associated kidney malformations

OBJECTIVES: ‘Persistent cloaca’ is a severe malformation affecting females in which the urinary, genital and alimentary tracts share a single conduit. Previously, a Uroplakin IIIA (UPIIIA) mutation was reported in one individual with persistent cloaca, and UPIIIA, Sonic Hedgehog (SHH), Ephrin B2 (EFNB2) and Hepatocyte Nuclear Factor 1β (HNF1β) are expressed during the normal development of organs that are affected in this condition. HNF1β mutations have been associated with uterine malformations in humans, and mutations of genes homologous to human SHH or EFNB2 cause persistent cloaca in mice. PATIENTS AND METHODS: We sought mutations of coding regions of UPIIIA, SHH, EFNB2 and HNF1β genes by direct sequencing in a group of 20 patients with persistent cloaca. Most had associated malformations of the upper renal tract and over half had impaired renal excretory function. The majority of patients had congenital anomalies outside the renal/genital tracts and two had the VACTERL association. RESULTS: Apart from a previously described index case, we failed to find UPIIIA mutations, and no patient had a SHH, EFNB2 or HNF1β mutation. CONCLUSION: Persistent cloaca is only rarely associated with UPIIIA mutation. Despite the fact that SHH and EFNB2 are appealing candidate genes, based on their expression patterns and mutant mice phenotypes, they were not mutated in these humans with persistent cloaca. Although HNF1β mutations can perturb paramesonephric duct fusion in humans, HNF1β was not mutated in persistent cloaca