Danielle Lellek Brooker Interview, April 23, 2021

In this interview, Danielle Brooker talks about her role as the general manager of Draught Works in Missoula, where she has worked since 2012. She discusses how Draught Works has a good relationship with the health department and how they have been adhering to their directives due to the COVID-19 pandemic. As a result of the directives, Brooker mentions how Draught Works implemented their changes to capacity, including using outdoor spaces, creating online ordering, and installing chip readers for contactless payment. As a result of following the health department’s directives, Brooker states that there were practically no cases of COVID-19 among the employees of Draught Works. Brooker discusses how they had just created a seltzer before the COVID-19 pandemic, which they were able to offer during the pandemic alongside their extensive beer list. Brooker also discusses how staff use social media to communicate with customers on various platforms.https://scholarworks.umt.edu/wetmissoulacovid19_oralhistory/1010/thumbnail.jp

The antiretroviral potency of APOBEC1 deaminase from small animal species

Although the role of the APOBEC3-dependent retroelement restriction system as an intrinsic immune defense against human immunodeficiency virus type1 (HIV-1) infection is becoming clear, only the rat ortholog of mammalian APOBEC1s (A1) thus far has been shown to possess antiviral activity. Here, we cloned A1 cDNAs from small animal species, and showed that similar to rat A1, both wild-type and Δvif HIV-1 infection was inhibited by mouse and hamster A1 (4- to 10-fold), whereas human A1 had negligible effects. Moreover, rabbit A1 significantly reduced the infectivity of both HIV-1 virions (>300-fold), as well as that of SIVmac, SIVagm, FIV and murine leukemia virus. Immunoblot analysis showed that A1s were efficiently incorporated into the HIV-1 virion, and their packaging is mediated through an interaction with the nucleocapsid Gag domain. Interestingly, there was a clear accumulation of particular C-T changes in the genomic RNAs of HIV-1 produced in their presence, with few G-A changes in the proviral DNA. Together, these data reveal that A1 may function as a defense mechanism, regulating retroelements in a wide range of mammalian species

A Survey of Genomic Traces Reveals a Common Sequencing Error, RNA Editing, and DNA Editing

While it is widely held that an organism's genomic information should remain constant, several protein families are known to modify it. Members of the AID/APOBEC protein family can deaminate DNA. Similarly, members of the ADAR family can deaminate RNA. Characterizing the scope of these events is challenging. Here we use large genomic data sets, such as the two billion sequences in the NCBI Trace Archive, to look for clusters of mismatches of the same type, which are a hallmark of editing events caused by APOBEC3 and ADAR. We align 603,249,815 traces from the NCBI trace archive to their reference genomes. In clusters of mismatches of increasing size, at least one systematic sequencing error dominates the results (G-to-A). It is still present in mismatches with 99% accuracy and only vanishes in mismatches at 99.99% accuracy or higher. The error appears to have entered into about 1% of the HapMap, possibly affecting other users that rely on this resource. Further investigation, using stringent quality thresholds, uncovers thousands of mismatch clusters with no apparent defects in their chromatograms. These traces provide the first reported candidates of endogenous DNA editing in human, further elucidating RNA editing in human and mouse and also revealing, for the first time, extensive RNA editing in Xenopus tropicalis. We show that the NCBI Trace Archive provides a valuable resource for the investigation of the phenomena of DNA and RNA editing, as well as setting the stage for a comprehensive mapping of editing events in large-scale genomic datasets

Mouse Apolipoprotein B Editing Complex 3 (APOBEC3) Is Expressed in Germ Cells and Interacts with Dead-End (DND1)

encoded protein, DND1, is able to bind to the 3′-untranslated region (UTR) of messenger RNAs (mRNAs) to displace micro-RNA (miRNA) interaction with mRNA. Thus, one function of DND1 is to prevent miRNA mediated repression of mRNA. We report that DND1 interacts specifically with APOBEC3. APOBEC3 is a multi-functional protein. It inhibits retroviral replication. In addition, recent studies show that APOBEC3 interacts with cellular RNA-binding proteins and to mRNA to inhibit miRNA-mediated repression of mRNA.Here we show that DND1 specifically interacts with another cellular protein, APOBEC3. We present our data which shows that DND1 co-immunoprecipitates APOBEC3 from mammalian cells and also endogenous APOBEC3 from mouse gonads. Whether the two proteins interact directly remains to be elucidated. We show that both DND1 and APOBEC3 are expressed in germ cells and in the early gonads of mouse embryo. Expression of fluorescently-tagged DND1 and APOBEC3 indicate they localize to the cytoplasm and when DND1 and APOBEC3 are expressed together in cells, they sequester near peri-nuclear sites.The 3′-UTR of mRNAs generally encode multiple miRNA binding sites as well as binding sites for a variety of RNA binding proteins. In light of our findings of DND1-APOBEC3 interaction and taking into consideration reports that DND1 and APOBEC3 bind to mRNA to inhibit miRNA mediated repression, our studies implicate a possible role of DND1-APOBEC3 interaction in modulating miRNA-mediated mRNA repression. The interaction of DND1 and APOBEC3 could be one mechanism for maintaining viability of germ cells and for preventing germ cell tumor development

APOBEC3G and APOBEC3F Require an Endogenous Cofactor to Block HIV-1 Replication

APOBEC3G (A3G)/APOBEC3F (A3F) are two members of APOBEC3 cytidine deaminase subfamily. Although they potently inhibit the replication of vif-deficient HIV-1, this mechanism is still poorly understood. Initially, A3G/A3F were thought to catalyze C-to-U transitions on the minus-strand viral cDNAs during reverse transcription to disrupt the viral life cycle. Recently, it was found more likely that A3G/A3F directly interrupts viral reverse transcription or integration. In addition, A3G/A3F are both found in the high-molecular-mass complex in immortalized cell lines, where they interact with a number of different cellular proteins. However, there has been no evidence to prove that these interactions are required for A3G/A3F function. Here, we studied A3G/A3F-restricted HIV-1 replication in six different human T cell lines by infecting them with wild-type or vif-deficient HIV-1. Interestingly, in a CEM-derived cell line CEM-T4, which expresses high levels of A3G/A3F proteins, the vif-deficient virus replicated as equally well as the wild-type virus, suggesting that these endogenous antiretroviral genes lost anti-HIV activities. It was confirmed that these A3G/A3F genes do not contain any mutation and are functionally normal. Consistently, overexpression of exogenous A3G/A3F in CEM-T4 cells still failed to restore their anti-HIV activities. However, this activity could be restored if CEM-T4 cells were fused to 293T cells to form heterokaryons. These results demonstrate that CEM-T4 cells lack a cellular cofactor, which is critical for A3G/A3F anti-HIV activity. We propose that a further study of this novel factor will provide another strategy for a complete understanding of the A3G/A3F antiretroviral mechanism

Produktivitätsmessung von Dienstleistungen – Entwicklung eines Messansatzes für die Produktivitätsbewertung von Dienstleistungen am Beispiel von Facility Services

Durch den zunehmenden Wettbewerbsdruck sehen sich Dienstleistungsunternehmen gezwungen ihre Leistungsproduktion zu professionalisieren und ressourcenschonend auszugestalten. Dieses Bestreben erfordert ein strukturiertes Dienstleistungsmanagement, welches durch kennzahlengestützte Steuerungsinstrumente bisher nicht ausgeschöpfte, versteckte Produktivitätspotenziale aufdeckt und zum Ausbau der betrieblichen Unternehmenssituation nutzt. Diese Notwendigkeit wird insbesondere im Business-to-Business-Bereich ersichtlich, wenn unternehmensbezogene Dienstleistungen ressourcenschonende Leistungen zu einem bestimmten Leistungsstandard hervorbringen müssen. Das Facility Management, welches die Steuerung und Koordination immobilienbezogener Dienstleistungen zur Unterstützung kerngeschäftlicher Aktivitäten umfasst, enthält als funktionales Subsystem derartige werttreibende Potenziale, die nach Freisetzung in die primären Wertaktivitäten einfließen können. Der Einsatz leistungsbasierter Instrumente zur Produktivitätsbewertung von Facility Services wird somit notwendig. Während im Sachgüterbereich die Produktivitätsmessung bereits ein prominentes Steuerungsinstrument darstellt, besteht im Dienstleistungsbereich bis dato ein wenig konsistentes Begriffsverständnis, sodass sich unterschiedliche Herausforderungen für die konzeptionelle Entwicklung eines Produktivitätsmesssystems ergeben. Vor diesem Hintergrund beruht das methodische Vorgehen der Modellentwicklung auf einer belastbaren, theoretischen Auseinandersetzung mit dem Status Quo des Produktivitätsbegriffs und der Harmonisierung dessen mit den Anforderungen der betrieblichen Praxis. Anhand einer qualitativen Literaturanalyse wurden relevante Produktivitätsfaktoren und damit verbundene Bewertungsindikatoren identifiziert, die anschließend in einem diskursiven Expertenworkshop hinsichtlich ihrer Validität und Anwendbarkeit bewertet und in ein konzeptionelles Messmodell überführt wurden. Basierend auf der klassischen Input-Output-Relation setzt sich die Produktivitätskennziffer im entwickelten Messmodell aus der Erhebung des Leistungsaufwands (Input) und des generierten Leistungsumsatzes (Output) zusammen, welche um die Qualitätsdimensionen des Leistungsergebnisses (Nachbesserungen bei Schlechtleistung, die den Gesamtleistungsaufwand verhältnisentsprechend erhöhen) ergänzt werden. Das entwickelte Messmodell wurde ebenfalls im Rahmen einer Fallstudienanalyse evaluiert und im Ergebnis als valides, praxistaugliches Instrument bewertet. Als generalistischer Messansatz, der auf einem monetären Bewertungsvorgehen beruht, lässt sich die Produktivitätsmessung somit auch auf andere Dienstleistungsbereiche übertragen. Im Ergebnis konnte anhand der dargestellten Untersuchungen das heterogene Begriffsverständnis über die Dienstleistungsproduktivität harmonisiert und ein Wissensbeitrag durch die Operationalisierung des Produktivitätsbegriffs, der als leistungsbasierte Produktivitätskennziffer in einen praxisnahen Messansatz eingebettet wurde, erzielt werden