THE commemoration was heartfelt, the nostalgia moving, the gratitude profound.

ROYAL CARIBBEAN has made further changes to their policy in a bid to instil confidence in customers and reduce the risk of financial losses for customers.

Shelves went tumbling down at Walmart.

F-35 jets fly over Utah as tribute to workers on frontline of COVID-19 fight

Teachers from North Elementary School in Noblesville decorated their cars and paraded through neighborhoods, waving and honking at students from afar during the closure of schools because of the coronavirus outbreak.

Officials would direct the Navy's Blue Angels and Air Force's Thunderbirds to fly over Indianapolis, Washington, New York and others

2020-03-27 -

Indiana will issue unemployment benefits to workers who do not typically qualify. But distributing new federal stimulus money will take time.

Indianapolis rapper Tevin Studdard celebrates the return of Long's Bakery with "Long's Bakery Music Video," a tribute to glazed yeast doughnuts.

The DOJ ordered an Orange County company to stop distributing hand-sanitizer products until it got FDA approval or removed certain claims from its labeling.

Saudi plans to build a new tech city are coming up against two hurdles: funding and a local tribe.

A chronology of key events

Key facts, figures and dates

Albert Jarrett was among some 10,000 West Indians who crossed the Atlantic to join the war effort.

1. After death, special trains take 16 migrant workers to home districts  The Tribune India
2. Unable to Overcome Impact Of Tragedy: Aurangabad Train Accident Survivor  NDTV
3. NHRC issues notice to Maharashtra govt over deaths of migrant workers in Aurangabad train mishap, seeks det...  Zee News
4. Tragedy on tracks  The Indian Express
5. Coronavirus Essential podcast | Aurangabad train mows down 17 migrant workers; COVID-19 spreads in 8 Maharashtra jails  Moneycontrol.com
6. View Full coverage on Google News

A game that punctures a hole in the noxious ideology that work gives meaning to our lives.

The Patriots are coming off a devastating 34-33 loss to the Miami Dolphins yet were lucky to maintain their playoff position as the No. 2 seed in the AFC

Laine Hardy and Maelyn Jarmon may have won different shows, but their experiences are very similar.

Octoshape announced a partnership with INVISO to deliver Internet TV contribution services throughout Latin America.

"At INVISO, we seek the most innovative and high quality products to serve our customers through the brands we represent,” said Jose Luis Reyes, Vice President for Sales and Operations, INVISO. “In the case of Octoshape, we found a company and a product that bring these qualities to our supply chain, sales and service.”

Octoshape offers an innovative cloud-based solution that provides instant infrastructure for the distribution of both linear and video on demand content. The Octoshape Infinite Uplink service provides point-to-point distribution of TV signals over the Internet for source signal acquisition to traditional IPTV and cable headends as an alternative to traditional methods like satellite and video fiber.

His unforced errors would make it hard on a U.S. manufacturer.

“It’s just so unfair that American companies aren’t allowed to pay bribes to get business overseas.”

oscon: RT @mmmpork: If you're going to #oscon and you're into #perl check out my talk on contributing http://t.co/HEJaPF5JbY

strataconf: As infrastructure gets simpler, interesting big data apps can emerge on BDAS, @Cloudera & other Hadoop distributions http://t.co/jaATalPL4P

Node.js is used by many companies for building performant backend services without sacrificing developer efficiency. In this hands-on guide, author Thomas Hunter II proves that Node.js is just as capable as traditional enterprise platforms for building services that are observable, scalable, and resilient. Intermediate to advanced Node.js developers will find themselves integrating application code with a breadth of tooling from each layer of a modern service stack.

Hear about travel to the Windward Islands of the Lesser Antilles in the Caribbean as the Amateur Traveler  talks again to Gary Arndt of Everything-Everyhwere.com. Gary just finished an island hopping tour that took him to most of the islands in the Caribbean. We will cover the islands of St. Martin, Anguilla, Saba, St. Barts, St. Eustatius, St. Kitts, Nevis, Antigua, Barbuda, and Montserrat on this episode.

Hear about travel to the Windward Islands of the Lesser Antilles in the Caribbean as the Amateur Traveler talks to Gary Arndt about his journey to visit many of the islands in the Caribbean on a single trip. In this episode we talk about Dominica, Martinique, Saint Lucia, Saint Vincent and the Grenadines, Grenada, Barbados, and Trinidad. This episode is a continuation of episode 386 which talked about the Leeward Islands.

Hear about travel to the countries of Kiribati, Tuvalu and Nauru in Micronesia as the Amateur Traveler talks to Stefan from Rapid Travel Chai about these small difficult to reach island nations. 

Interferon-regulated myxovirus resistance protein B (MxB) is an interferon-induced GTPase belonging to the dynamin superfamily. It inhibits infection with a wide range of different viruses, including HIV-1, by impairing viral DNA entry into the nucleus. Unlike the related antiviral GTPase MxA, MxB possesses an N-terminal region that contains a nuclear localization signal and is crucial for inhibiting HIV-1. Because MxB previously has been shown to reside in both the nuclear envelope and the cytoplasm, here we used bioinformatics and biochemical approaches to identify a nuclear export signal (NES) responsible for MxB's cytoplasmic location. Using the online computational tool LocNES (Locating Nuclear Export Signals or NESs), we identified five putative NES candidates in MxB and investigated whether their deletion caused nuclear localization of MxB. Our results revealed that none of the five deletion variants relocates to the nucleus, suggesting that these five predicted NES sequences do not confer NES activity. Interestingly, deletion of one sequence, encompassing amino acids 505–527, abrogated the anti-HIV-1 activity of MxB. Further mutation experiments disclosed that amino acids 515–519, and Pro-515 in particular, regulate MxB oligomerization and its binding to HIV-1 capsid, thereby playing an important role in MxB-mediated restriction of HIV-1 infection. In summary, our results indicate that none of the five predicted NES sequences in MxB appears to be required for its nuclear export. Our findings also reveal several residues in MxB, including Pro-515, critical for its oligomerization and anti-HIV-1 function.

The formation of translationally inactive 70S dimers (called 100S ribosomes) by hibernation-promoting factor is a widespread survival strategy among bacteria. Ribosome dimerization is thought to be reversible, with the dissociation of the 100S complexes enabling ribosome recycling for participation in new rounds of translation. The precise pathway of 100S ribosome recycling has been unclear. We previously found that the heat-shock GTPase HflX in the human pathogen Staphylococcus aureus is a minor disassembly factor. Cells lacking hflX do not accumulate 100S ribosomes unless they are subjected to heat exposure, suggesting the existence of an alternative pathway during nonstressed conditions. Here, we provide biochemical and genetic evidence that two essential translation factors, ribosome-recycling factor (RRF) and GTPase elongation factor G (EF-G), synergistically split 100S ribosomes in a GTP-dependent but tRNA translocation-independent manner. We found that although HflX and the RRF/EF-G pair are functionally interchangeable, HflX is expressed at low levels and is dispensable under normal growth conditions. The bacterial RRF/EF-G pair was previously known to target only the post-termination 70S complexes; our results reveal a new role in the reversal of ribosome hibernation that is intimately linked to bacterial pathogenesis, persister formation, stress responses, and ribosome integrity.

Interferon-regulated myxovirus resistance protein B (MxB) is an interferon-induced GTPase belonging to the dynamin superfamily. It inhibits infection with a wide range of different viruses, including HIV-1, by impairing viral DNA entry into the nucleus. Unlike the related antiviral GTPase MxA, MxB possesses an N-terminal region that contains a nuclear localization signal and is crucial for inhibiting HIV-1. Because MxB previously has been shown to reside in both the nuclear envelope and the cytoplasm, here we used bioinformatics and biochemical approaches to identify a nuclear export signal (NES) responsible for MxB's cytoplasmic location. Using the online computational tool LocNES (Locating Nuclear Export Signals or NESs), we identified five putative NES candidates in MxB and investigated whether their deletion caused nuclear localization of MxB. Our results revealed that none of the five deletion variants relocates to the nucleus, suggesting that these five predicted NES sequences do not confer NES activity. Interestingly, deletion of one sequence, encompassing amino acids 505–527, abrogated the anti-HIV-1 activity of MxB. Further mutation experiments disclosed that amino acids 515–519, and Pro-515 in particular, regulate MxB oligomerization and its binding to HIV-1 capsid, thereby playing an important role in MxB-mediated restriction of HIV-1 infection. In summary, our results indicate that none of the five predicted NES sequences in MxB appears to be required for its nuclear export. Our findings also reveal several residues in MxB, including Pro-515, critical for its oligomerization and anti-HIV-1 function.

The formation of translationally inactive 70S dimers (called 100S ribosomes) by hibernation-promoting factor is a widespread survival strategy among bacteria. Ribosome dimerization is thought to be reversible, with the dissociation of the 100S complexes enabling ribosome recycling for participation in new rounds of translation. The precise pathway of 100S ribosome recycling has been unclear. We previously found that the heat-shock GTPase HflX in the human pathogen Staphylococcus aureus is a minor disassembly factor. Cells lacking hflX do not accumulate 100S ribosomes unless they are subjected to heat exposure, suggesting the existence of an alternative pathway during nonstressed conditions. Here, we provide biochemical and genetic evidence that two essential translation factors, ribosome-recycling factor (RRF) and GTPase elongation factor G (EF-G), synergistically split 100S ribosomes in a GTP-dependent but tRNA translocation-independent manner. We found that although HflX and the RRF/EF-G pair are functionally interchangeable, HflX is expressed at low levels and is dispensable under normal growth conditions. The bacterial RRF/EF-G pair was previously known to target only the post-termination 70S complexes; our results reveal a new role in the reversal of ribosome hibernation that is intimately linked to bacterial pathogenesis, persister formation, stress responses, and ribosome integrity.

Heme-regulatory motifs (HRMs) are present in many proteins that are involved in diverse biological functions. The C-terminal tail region of human heme oxygenase-2 (HO2) contains two HRMs whose cysteine residues form a disulfide bond; when reduced, these cysteines are available to bind Fe3+-heme. Heme binding to the HRMs occurs independently of the HO2 catalytic active site in the core of the protein, where heme binds with high affinity and is degraded to biliverdin. Here, we describe the reversible, protein-mediated transfer of heme between the HRMs and the HO2 core. Using hydrogen-deuterium exchange (HDX)-MS to monitor the dynamics of HO2 with and without Fe3+-heme bound to the HRMs and to the core, we detected conformational changes in the catalytic core only in one state of the catalytic cycle—when Fe3+-heme is bound to the HRMs and the core is in the apo state. These conformational changes were consistent with transfer of heme between binding sites. Indeed, we observed that HRM-bound Fe3+-heme is transferred to the apo-core either upon independent expression of the core and of a construct spanning the HRM-containing tail or after a single turnover of heme at the core. Moreover, we observed transfer of heme from the core to the HRMs and equilibration of heme between the core and HRMs. We therefore propose an Fe3+-heme transfer model in which HRM-bound heme is readily transferred to the catalytic site for degradation to facilitate turnover but can also equilibrate between the sites to maintain heme homeostasis.

Incorporation of ribonucleotides into DNA can severely diminish genome integrity. However, how ribonucleotides instigate DNA damage is poorly understood. In DNA, they can promote replication stress and genomic instability and have been implicated in several diseases. We report here the impact of the ribonucleotide rATP and of its naturally occurring damaged analog 1,N6-ethenoadenosine (1,N6-ϵrA) on translesion synthesis (TLS), mediated by human DNA polymerase η (hpol η), and on RNase H2–mediated incision. Mass spectral analysis revealed that 1,N6-ϵrA in DNA generates extensive frameshifts during TLS, which can lead to genomic instability. Moreover, steady-state kinetic analysis of the TLS process indicated that deoxypurines (i.e. dATP and dGTP) are inserted predominantly opposite 1,N6-ϵrA. We also show that hpol η acts as a reverse transcriptase in the presence of damaged ribonucleotide 1,N6-ϵrA but has poor RNA primer extension activities. Steady-state kinetic analysis of reverse transcription and RNA primer extension showed that hpol η favors the addition of dATP and dGTP opposite 1,N6-ϵrA. We also found that RNase H2 recognizes 1,N6-ϵrA but has limited incision activity across from this lesion, which can lead to the persistence of this detrimental DNA adduct. We conclude that the damaged and unrepaired ribonucleotide 1,N6-ϵrA in DNA exhibits mutagenic potential and can also alter the reading frame in an mRNA transcript because 1,N6-ϵrA is incompletely incised by RNase H2.

The ribosome biogenesis factor Las1 is an essential endoribonuclease that is well-conserved across eukaryotes and a newly established member of the higher eukaryotes and prokaryotes nucleotide-binding (HEPN) domain-containing nuclease family. HEPN nucleases participate in diverse RNA cleavage pathways and share a short HEPN nuclease motif (RφXXXH) important for RNA cleavage. Most HEPN nucleases participate in stress-activated RNA cleavage pathways; Las1 plays a fundamental role in processing pre-rRNA. Underscoring the significance of Las1 function in the cell, mutations in the human LAS1L (LAS1-like) gene have been associated with neurological dysfunction. Two juxtaposed HEPN nuclease motifs create Las1's composite nuclease active site, but the roles of the individual HEPN motif residues are poorly defined. Here using a combination of in vivo experiments in Saccharomyces cerevisiae and in vitro assays, we show that both HEPN nuclease motifs are required for Las1 nuclease activity and fidelity. Through in-depth sequence analysis and systematic mutagenesis, we determined the consensus HEPN motif in the Las1 subfamily and uncovered its canonical and specialized elements. Using reconstituted Las1 HEPN-HEPN' chimeras, we defined the molecular requirements for RNA cleavage. Intriguingly, both copies of the Las1 HEPN motif were important for nuclease function, revealing that both HEPN motifs participate in coordinating the RNA within the Las1 active site. We also established that conformational flexibility of the two HEPN domains is important for proper nuclease function. The results of our work reveal critical information about how dual HEPN domains come together to drive Las1-mediated RNA cleavage.

19 December 2013

The ribosome biogenesis factor Las1 is an essential endoribonuclease that is well-conserved across eukaryotes and a newly established member of the higher eukaryotes and prokaryotes nucleotide-binding (HEPN) domain-containing nuclease family. HEPN nucleases participate in diverse RNA cleavage pathways and share a short HEPN nuclease motif (RφXXXH) important for RNA cleavage. Most HEPN nucleases participate in stress-activated RNA cleavage pathways; Las1 plays a fundamental role in processing pre-rRNA. Underscoring the significance of Las1 function in the cell, mutations in the human LAS1L (LAS1-like) gene have been associated with neurological dysfunction. Two juxtaposed HEPN nuclease motifs create Las1's composite nuclease active site, but the roles of the individual HEPN motif residues are poorly defined. Here using a combination of in vivo experiments in Saccharomyces cerevisiae and in vitro assays, we show that both HEPN nuclease motifs are required for Las1 nuclease activity and fidelity. Through in-depth sequence analysis and systematic mutagenesis, we determined the consensus HEPN motif in the Las1 subfamily and uncovered its canonical and specialized elements. Using reconstituted Las1 HEPN-HEPN' chimeras, we defined the molecular requirements for RNA cleavage. Intriguingly, both copies of the Las1 HEPN motif were important for nuclease function, revealing that both HEPN motifs participate in coordinating the RNA within the Las1 active site. We also established that conformational flexibility of the two HEPN domains is important for proper nuclease function. The results of our work reveal critical information about how dual HEPN domains come together to drive Las1-mediated RNA cleavage.

8 April 2020

Imagine trying to describe the circulation and temperatures across the vast expanse of our oceans. Good models of our oceans not only benefit fishermen on our coasts but farmers inland as well. Until recently, there were neither adequate tools nor enough data to construct models. Now with new data and new mathematics, short-range climate forecasting for example, of an upcoming El Nino is possible.There is still much work to be done in long-term climate forecasting, however, and we only barely understand the oceans. Existing equations describe ocean dynamics, but solutions to the equations are currently out of reach. No computer can accommodate the data required to approximate a good solution to these equations. Researchers therefore make simplifying assumptions in order to solve the equations. New data are used to test the accuracy of models derived from these assumptions. This research is essential because we cannot understand our climate until we understand the oceans. For More Information: What.s Happening in the Mathematical Sciences, Vol 1, Barry Cipra.