A. Seesanea and I. E. Verbitsky
St. Petersburg Math. J. 31 (2020), 557-572.
Abstract, references and article information

Véronique Bazier-Matte and Pierre-Guy Plamondon
Proc. Amer. Math. Soc. 148 (2020), 2397-2409.
Abstract, references and article information

To celebrate the 30th anniversary of the Cultural Centre, the Leisure & Cultural Services Department will hold a fun day from noon to 5.30pm on November 9.

The centre’s venue partners, the Hong Kong Chinese Orchestra, the Hong Kong Philharmonic Orchestra, the Hong Kong Ballet and Zuni Icosahedron will use the latest technology in the foyer to present Western and Chinese music, a ballet performance and sound and scene recreation of the former Kowloon-Canton Railway Station.

At the piazza areas, more than 40 dancers will perform works by acclaimed and emerging choreographers and lead visitors to discover every corner of the centre.

Artist Enoch Cheng will curate "Unseen Scene" in the backstage area to display the magic behind a show through music, dance, projections and other special performances.

Additional programmes will include the Stage & Technology Workshop and the 30th anniversary exhibition "Traces of the Past & Future".

Besides free events on the fun day, the centre will hold a celebratory concert at 8pm on November 29 and 30. Pieces specially selected from the repertoire of the centre's opening concert 30 years ago will be presented.

Click here for details.

Asian Ethnic Cultural Performances 2019 will be staged on Sunday to demonstrate the diversity of Asia’s cultures.

The event will feature ethnic performances and activities with representatives from 19 Asian countries and places taking part.

Korean traditional Nongak and Taepyeongmu dances, a Japanese Tokushima Awa dance, Indian classical and Bollywood dances, and folk dance performances of countries such as Bangladesh, Laos, Vietnam, the Philippines and Sri Lanka will be showcased.

Hong Kong and Macau arts groups will stage a hip hop lion dance and a cheerleading performance.

Other activities include traditional arts and crafts, costume and food displays, as well as a bamboo musical instrument workshop.

A CIBS mobile studio supported by Radio Television Hong Kong will also be set up to let members of the public learn about broadcasting.

The event will be held from 2pm to 6pm at the Cultural Centre Piazza. Admission is free.

Click here for details.

The East Kowloon Cultural Centre Public Artwork Commissioning Project is inviting artists, designers and architects to create an art landscape for the centre.

Presented by the Leisure & Cultural Services Department and organised by the Art Promotion Office, the project endeavors to capture and showcase the past, present and future of East Kowloon.

The commissioned artworks will represent five themes - memory, impression, moment, dream and imagination - aiming to display the district’s changing nature, and to manifest the local community’s cultural sustainability, aspirations and dreams.

Interested participants must submit their preliminary proposal with their curriculum vitae, documentation of their previous artwork and the artistic concept of the proposed artwork.

All proposals must be delivered to the centre’s Public Art Project Management Team at G/F, 50-54 Lok Ku Road, Sheung Wan before 7pm on May 4.

Participants shortlisted by the selection panel are required to submit detailed proposals before 7pm on June 30 for the next stage of the selection process.

Up to three proposals shall be chosen by the selection panel for commissioning and installing in the centre.

Toni Tripp-Reimer
Jan 1, 2001; 14:
Articles

Myostatin (or growth/differentiation factor 8 (GDF8)) is a member of the transforming growth factor β superfamily of growth factors and negatively regulates skeletal muscle growth. Its dysregulation is implicated in muscle wasting diseases. SRK-015 is a clinical-stage mAb that prevents extracellular proteolytic activation of pro- and latent myostatin. Here we used integrated structural and biochemical approaches to elucidate the molecular mechanism of antibody-mediated neutralization of pro-myostatin activation. The crystal structure of pro-myostatin in complex with 29H4-16 Fab, a high-affinity variant of SRK-015, at 2.79 Å resolution revealed that the antibody binds to a conformational epitope in the arm region of the prodomain distant from the proteolytic cleavage sites. This epitope is highly sequence-divergent, having only limited similarity to other closely related members of the transforming growth factor β superfamily. Hydrogen/deuterium exchange MS experiments indicated that antibody binding induces conformational changes in pro- and latent myostatin that span the arm region, the loops contiguous to the protease cleavage sites, and the latency-associated structural elements. Moreover, negative-stain EM with full-length antibodies disclosed a stable, ring-like antigen–antibody structure in which the two Fab arms of a single antibody occupy the two arm regions of the prodomain in the pro- and latent myostatin homodimers, suggesting a 1:1 (antibody:myostatin homodimer) binding stoichiometry. These results suggest that SRK-015 binding stabilizes the latent conformation and limits the accessibility of protease cleavage sites within the prodomain. These findings shed light on approaches that specifically block the extracellular activation of growth factors by targeting their precursor forms.

Pyruvate kinase muscle isoform 2 (PKM2) is a key glycolytic enzyme involved in ATP generation and critical for cancer metabolism. PKM2 is expressed in many human cancers and is regulated by complex mechanisms that promote tumor growth and proliferation. Therefore, it is considered an attractive therapeutic target for modulating tumor metabolism. Various stimuli allosterically regulate PKM2 by cycling it between highly active and less active states. Several small molecules activate PKM2 by binding to its intersubunit interface. Serine and cysteine serve as an activator and inhibitor of PKM2, respectively, by binding to its amino acid (AA)-binding pocket, which therefore represents a potential druggable site. Despite binding similarly to PKM2, how cysteine and serine differentially regulate this enzyme remains elusive. Using kinetic analyses, fluorescence binding, X-ray crystallography, and gel filtration experiments with asparagine, aspartate, and valine as PKM2 ligands, we examined whether the differences in the side-chain polarity of these AAs trigger distinct allosteric responses in PKM2. We found that Asn (polar) and Asp (charged) activate PKM2 and that Val (hydrophobic) inhibits it. The results also indicate that both Asn and Asp can restore the activity of Val-inhibited PKM2. AA-bound crystal structures of PKM2 displayed distinctive interactions within the binding pocket, causing unique allosteric effects in the enzyme. These structure-function analyses of AA-mediated PKM2 regulation shed light on the chemical requirements in the development of mechanism-based small-molecule modulators targeting the AA-binding pocket of PKM2 and provide broader insights into the regulatory mechanisms of complex allosteric enzymes.

The C-type lectin receptors (CLRs) form a family of pattern recognition receptors that recognize numerous pathogens, such as bacteria and fungi, and trigger innate immune responses. The extracellular carbohydrate-recognition domain (CRD) of CLRs forms a globular structure that can coordinate a Ca2+ ion, allowing receptor interactions with sugar-containing ligands. Although well-conserved, the CRD fold can also display differences that directly affect the specificity of the receptors for their ligands. Here, we report crystal structures at 1.8–2.3 Å resolutions of the CRD of murine dendritic cell-immunoactivating receptor (DCAR, or Clec4b1), the CLR that binds phosphoglycolipids such as acylated phosphatidyl-myo-inositol mannosides (AcPIMs) of mycobacteria. Using mutagenesis analysis, we identified critical residues, Ala136 and Gln198, on the surface surrounding the ligand-binding site of DCAR, as well as an atypical Ca2+-binding motif (Glu-Pro-Ser/EPS168–170). By chemically synthesizing a water-soluble ligand analog, inositol-monophosphate dimannose (IPM2), we confirmed the direct interaction of DCAR with the polar moiety of AcPIMs by biolayer interferometry and co-crystallization approaches. We also observed a hydrophobic groove extending from the ligand-binding site that is in a suitable position to interact with the lipid portion of whole AcPIMs. These results suggest that the hydroxyl group-binding ability and hydrophobic groove of DCAR mediate its specific binding to pathogen-derived phosphoglycolipids such as mycobacterial AcPIMs.

Cell-surface signaling (CSS) in Gram-negative bacteria involves highly conserved regulatory pathways that optimize gene expression by transducing extracellular environmental signals to the cytoplasm via inner-membrane sigma regulators. The molecular details of ferric siderophore-mediated activation of the iron import machinery through a sigma regulator are unclear. Here, we present the 1.56 Å resolution structure of the periplasmic complex of the C-terminal CSS domain (CCSSD) of PupR, the sigma regulator in the Pseudomonas capeferrum pseudobactin BN7/8 transport system, and the N-terminal signaling domain (NTSD) of PupB, an outer-membrane TonB-dependent transducer. The structure revealed that the CCSSD consists of two subdomains: a juxta-membrane subdomain, which has a novel all-β-fold, followed by a secretin/TonB, short N-terminal subdomain at the C terminus of the CCSSD, a previously unobserved topological arrangement of this domain. Using affinity pulldown assays, isothermal titration calorimetry, and thermal denaturation CD spectroscopy, we show that both subdomains are required for binding the NTSD with micromolar affinity and that NTSD binding improves CCSSD stability. Our findings prompt us to present a revised model of CSS wherein the CCSSD:NTSD complex forms prior to ferric-siderophore binding. Upon siderophore binding, conformational changes in the CCSSD enable regulated intramembrane proteolysis of the sigma regulator, ultimately resulting in transcriptional regulation.

In cyanobacteria, metabolic pathways that use the nitrogen-rich amino acid arginine play a pivotal role in nitrogen storage and mobilization. The N-terminal domains of two recently identified bacterial enzymes: ArgZ from Synechocystis and AgrE from Anabaena, have been found to contain an arginine dihydrolase. This enzyme provides catabolic activity that converts arginine to ornithine, resulting in concomitant release of CO2 and ammonia. In Synechocystis, the ArgZ-mediated ornithine–ammonia cycle plays a central role in nitrogen storage and remobilization. The C-terminal domain of AgrE contains an ornithine cyclodeaminase responsible for the formation of proline from ornithine and ammonia production, indicating that AgrE is a bifunctional enzyme catalyzing two sequential reactions in arginine catabolism. Here, the crystal structures of AgrE in three different ligation states revealed that it has a tetrameric conformation, possesses a binding site for the arginine dihydrolase substrate l-arginine and product l-ornithine, and contains a binding site for the coenzyme NAD(H) required for ornithine cyclodeaminase activity. Structure–function analyses indicated that the structure and catalytic mechanism of arginine dihydrolase in AgrE are highly homologous with those of a known bacterial arginine hydrolase. We found that in addition to other active-site residues, Asn-71 is essential for AgrE's dihydrolase activity. Further analysis suggested the presence of a passage for substrate channeling between the two distinct AgrE active sites, which are situated ∼45 Å apart. These results provide structural and functional insights into the bifunctional arginine dihydrolase–ornithine cyclodeaminase enzyme AgrE required for arginine catabolism in Anabaena.

Knowledge of the molecular events in mitochondrial DNA (mtDNA) replication is crucial to understanding the origins of human disorders arising from mitochondrial dysfunction. Twinkle helicase is an essential component of mtDNA replication. Here, we employed atomic force microscopy imaging in air and liquids to visualize ring assembly, DNA binding, and unwinding activity of individual Twinkle hexamers at the single-molecule level. We observed that the Twinkle subunits self-assemble into hexamers and higher-order complexes that can switch between open and closed-ring configurations in the absence of DNA. Our analyses helped visualize Twinkle loading onto and unloading from DNA in an open-ringed configuration. They also revealed that closed-ring conformers bind and unwind several hundred base pairs of duplex DNA at an average rate of ∼240 bp/min. We found that the addition of mitochondrial single-stranded (ss) DNA–binding protein both influences the ways Twinkle loads onto defined DNA substrates and stabilizes the unwound ssDNA product, resulting in a ∼5-fold stimulation of the apparent DNA-unwinding rate. Mitochondrial ssDNA-binding protein also increased the estimated translocation processivity from 1750 to >9000 bp before helicase disassociation, suggesting that more than half of the mitochondrial genome could be unwound by Twinkle during a single DNA-binding event. The strategies used in this work provide a new platform to examine Twinkle disease variants and the core mtDNA replication machinery. They also offer an enhanced framework to investigate molecular mechanisms underlying deletion and depletion of the mitochondrial genome as observed in mitochondrial diseases.

Mozart once called the pipe organ the “king of instruments” and Hong Kong has musical royalty sitting in the heart of Tsim Sha Tsui.

It is hard to miss the pipe organ when you are inside the Cultural Centre Concert Hall. With its four manuals, 93 stops and 8,000 pipes, it can produce an exceptional array of timbre and volume.

“The pipe organ is the ‘pearl’ of a concert hall,” explained pipe organist Chiu Siu-ling.

“Its enormous range of sounds, coupled with a huge variety of tone colours, creates a remarkable musical experience equivalent to that of an orchestra. It is the king of instruments.”

Star performer

The four-storey tall pipe organ was hand-made by famed Austrian manufacturer Rieger Orgelbau and installed when the Cultural Centre was built 30 years ago. It has attracted several world-renowned performers to Hong Kong over the years.

“When we built it in 1989, it was the first, huge mechanically operated organ in Asia. I think it is still our biggest organ we have ever built in Asia,” Rieger pipe organ maintenance technician Gerhard Pohl said.

He added that the instrument is also remarkably versatile.

“There are many organs they are built just for one musical style. On this organ, you can play everything, even jazz music.”

Mr Pohl comes to Hong Kong once a year to give the 30-year-old organ its annual health check. In between visits by the manufacturer’s technicians, William Wen ensures the Cultural Centre’s star performer remains in tip-top condition.

Top tuner

Mr Wen is Hong Kong’s only pipe organ maintenance technician. He carries out routine checks on the instrument and tunes it every month.

“Keeping it in tune is my responsibility. I take care of it.”

Mr Wen was one of the first in Hong Kong to learn the instrument in the 1980s. He became interested in pipe organ maintenance after a visit to an organ manufacturing company in Europe.

He recalled that in the ‘80s, organists in Hong Kong were few and far between. He was called upon to test the Cultural Centre organ before the grand opening, making him one of the first organists to play it.

“It was dark in the hall, so at first, I only noticed the organ’s keys. When I got closer, I was astounded by just how big the instrument was.”

Mr Wen takes up to five hours per maintenance visit to thoroughly check the pipes and tune the organ. He explained that the organ is regularly updated and was fitted with an electrical coupler system, providing greater flexibility in switching between lighter and stronger key actions as desired by the player.

“It has been updated a few times. In 2010, the organ was installed with an electrical coupler system. I used to get someone to press the keys for me while I would tune it from inside. But now, I just need a mobile phone on which I can tune it by myself.”

Musical legacy

To promote organ music and nurture local organists, the Cultural Centre has been organising the “King of the Instruments” Pipe Organ Education Series annually since 1999. Now the programme’s principal instructor, Ms Chiu has been teaching from the outset.

“People are not allowed to touch the instrument in a lot of other concert halls, and they seldom hold pipe organ concerts. This is actually not good for the organ. It has to be played to keep it in good condition or the keys and parts will become hard and old. This inspires me teach and let more and more musicians learn and play the instrument,” Ms Chiu explained.

More than 300 organists have so far been trained on the Cultural Centre’s organ. But it has also played something other than music - it has played Cupid too.

Like-minded musicians

Pipe organist Simon Chan met his wife, Shirley Cheng, through the programme.

“I joined the Pipe Organ Education Series in 2000. After that, I continued learning the organ with Ms Chiu and eventually became one of the course tutors. I met my wife in 2003 on the course.”

Ms Cheng was also a student on the course. She switched majors from information technology to music and that fateful decision led to meeting her husband.

“He asked me to watch him play. He was playing a big piece which was powerful and loud, and that was when I fell for his charms.”

The couple decided to embark on a life journey together and now hope to pass on their love of music and the pipe organ to their children.

“I wish for my kids to learn music too and to find out more about the pipe organ. This instrument brought their father and mother together and I want them to know our story,” said Ms Cheng.

The organist-music teacher duo also hopes to share the joy of the pipe organ with the rest of the community.

(American Friends of Tel Aviv University) The research reveals that hunter-gatherer societies expressed a deep emotional and psychological connection with the animal species they hunted, especially after their disappearance. The study will help anthropologists and others understand the profound environmental changes taking place in our own lifetimes.

(PLOS) The ancient cemetery of Mözs-Icsei d?l? in present-day Hungary holds clues to a unique community formation during the beginnings of Europe's Migration Period, according to a study published April 29, 2020 in the open-access journal PLOS ONE by Corina Knipper from the Curt-Engelhorn-Center for Archaeometry, Germany, István Koncz, Tivadar Vida from the Eötvös Loránd University, Budapest, Hungary and colleagues.

Toni Tripp-Reimer
Jan 1, 2001; 14:
Articles

Elena Lorente
Apr 7, 2020; 0:RA120.002014v1-mcp.RA120.002014
Research