Acetylation has recently emerged as an important mechanism for controlling a broad array of proteins mediating cellular adaptation to metabolic fuels. is governed, in part, by SIRTs (sirtuins), class III NAD+-dependent deacetylases that regulate lipid and glucose metabolism liver during fasting aging. However, the role acetylation or pathogenic hepatic fuel under nutrient excess unknown. In present study, we isolated acetylated from total proteome observed 193 preferentially mice fed on HFD...

Intestinal epithelial cells (IECs) are exposed to profound fluctuations in oxygen tension and have evolved adaptive transcriptional responses a low-oxygen environment. These adaptations mediated primarily through the hypoxia-inducible factor (HIF) complex. Given central role of IEC barrier function, we sought determine whether HIF influenced tight junction (TJ) structure function. Initial studies revealed that short hairpin RNA-mediated depletion HIF1β T84 resulted defects nonuniform,...

Significance Intestinal epithelial barrier dysregulation is a hallmark of inflammatory bowel diseases (IBDs). A central role for hypoxic signaling has been defined in modulation during inflammation. We demonstrate that genes involved creatine metabolism, the kinases (CKs), are coordinately regulated by hypoxia-inducible transcription factors (HIFs) and such regulation critical to function. Inhibition CK pathway abrogates apical junction assembly integrity. Dietary supplementation profoundly...

The unidirectional and opposite-polarity microtubule-based motors, dynein kinesin, drive long-distance intracellular cargo transport. Cellular observations suggest that motors may be coupled. We recently identified an interaction between the cytoplasmic dynein-1 activating adaptor Hook3 kinesin-3 KIF1C. Here, using in vitro reconstitutions with purified components, we show KIF1C dynein/dynactin can exist a complex scaffolded by Hook3. Full-length binds to activates motility. also short...

In eukaryotic cells, intracellular components are organized by the microtubule motors cytoplasmic dynein-1 (dynein) and kinesins, which linked to cargos via adaptor proteins. While ~40 kinesins transport cargo toward plus end of microtubules, a single dynein moves in opposite direction. How transports wide variety remains an open question. The FTS-Hook-FHIP ('FHF') complex links humans fungi. As human cells have three Hooks four FHIP proteins, we hypothesized that combinatorial assembly...

Abstract Cytoplasmic dynein-1 transports intracellular cargo towards microtubule minus ends. Dynein is autoinhibited and undergoes conformational changes to form an active complex that consists of one or two dynein dimers, the dynactin complex, activating adapter(s). The Lissencephaly 1 gene, LIS1 , genetically linked pathway from fungi mammals mutated in people with neurodevelopmental disease lissencephaly. Lis1 required for complexes form, but how it enables this unclear. Here, we present...

Extracellular vesicles (EVs) are key contributors to cancer where they play an integral role in cell-cell communication and transfer pro-oncogenic molecules recipient cells thereby conferring a cancerous phenotype. Here, we purified EVs using straightforward biochemical approaches from multiple cell lines subsequently characterized these via biophysical methods. In addition, used fluorescence microscopy directly show internalization of into the within few minutes upon addition cells. We...

// Agnieszka A. Kendrick 1 , Johnathon Schafer Monika Dzieciatkowska Travis Nemkov Angelo D’Alessandro Deepika Neelakantan 2 Heide L. Ford Chad G. Pearson 3 Colin D. Weekes 4 Kirk C. Hansen Elan Z. Eisenmesser Department of Biochemistry and Molecular Genetics, School Medicine, University Colorado Denver, CO, USA Pharmacology, Cell Developmental Biology, Division Oncology, Correspondence to: Eisenmesser, email: elan.eisenmesser@ucdenver.edu Keywords: metabolism, ancillary protein, PDAC,...

Cytoplasmic dynein-1 (dynein) is an essential molecular motor in eukaryotic cells. Dynein primarily exists autoinhibited Phi state and requires conformational changes to assemble with its cofactors form active transport complexes. LIS1, a key dynein regulator, enhances activation assembly. Using cryo-EM human dynein-LIS1 sample incubated ATP, we mapped the landscape of by LIS1. We discovered novel early intermediate that propose precedes our previously identified Chi state. Mutations disrupt...

Interleukin-8 (CXCL8, IL-8) is a proinflammatory chemokine important for the regulation of inflammatory and immune responses via its interaction with G-protein coupled receptors, including CXC receptor 1 (CXCR1). CXCL8 exists as both monomer dimer at physiological concentrations, yet molecular basis well importance formation remain poorly characterized. Although several biological studies have indicated that are active, biophysical reported conflicting results regarding binding to CXCR1. To...

IgA1 proteases (IgA1P) from diverse pathogenic bacteria specifically cleave human immunoglobulin A1 (IgA1) at the hinge region, thereby thwarting protective host immune responses. Streptococcus pneumoniae (S. pneumoniae) IgA1P shares no sequence conservation with serine or cysteine types of IgA1Ps other known proteins, than a conserved HExxH Zn-binding motif (1604-1608) found in metalloproteases. We have developed novel expression system to produce mature S. and we discovered that this form...

Most organelles move bidirectionally on microtubule tracks, yet how this opposing movement is regulated by kinesin and dynein remains unclear. Recent work found that ARL8, a known anterograde adaptor linking the lysosome to kinesin, also links lysosomes retrograde motor dynein, providing key insight into bidirectional organelle in cells.

The coronavirus disease 2019 (COVID-19) pandemic has had a detrimental effect on research. However, little been done to identify and solve the unique challenges faced by early career investigators (ECIs). As group of American Cancer Society-funded ECIs, we provide recommendations for solving these in aftermath pandemic. COVID-19 resulted stressors affecting including graduate students postdoctoral fellows (ECI-trainees), assistant professors (ECI-faculty). Some include heavier workloads, job...

Cytoplasmic dynein-1 transports many intracellular cargos towards microtubule minus ends. Dynein is autoinhibited and undergoes conformational changes to form an active complex, consisting of one or two dynein dimers, the dynactin complex activating adaptor(s) 1,2 . The Lissencephaly 1 gene, LIS1 , genetically linked pathway from fungi mammals mutated in patients with neurodevelopmental disease lissencephaly 3–5 Lis1 required for complexes 6–10 but how it does so unclear. Here, we present a...

Abstract The unidirectional and opposite-polarity microtubule-based motors, dynein kinesin, drive long-distance intracellular cargo transport. Cellular observations support the existence of mechanisms to couple opposite polarity motors: in cells some cargos rapidly switch directions kinesin motors can be used localize dynein. We recently identified an interaction between cytoplasmic dynein-1 activating adaptor HOOK3 kinesin-3 KIF1C. Here we show that KIF1C dynein/dynactin exist a single...

Cytoplasmic dynein-1 (dynein) is an essential molecular motor controlled in part by autoinhibition. We recently identified a structure of partially autoinhibited dynein bound to Lis1, key regulator mutated the neurodevelopmental disease lissencephaly. This provides intermediate state activation pathway; however, other structural information needed fully explain Lis1 function activation. Here, we used cryo-EM and samples incubated with ATP for different times reveal novel conformations that...

The gastrointestinal mucosa provides the primary physical barrier against gut luminal contents. In mucosal diseases such as inflammatory bowel disease, hypoxia and inflammation occur coincidentally at level of epithelium. Adaptive transcriptional responses to oxygen deprivation are mediated primarily through hypoxia‐inducible factor complex (HIF), comprised an oxygen‐labile ‘ Α’ subunit constitutively expressed ‘Β’ subunit. A role for HIF has been implicated in orchestrating a signaling...

Abstract In eukaryotic cells, intracellular components are organized by the microtubule motors cytoplasmic dynein-1 (dynein) and kinesins, which linked to cargos via adaptor proteins. While ∼40 kinesins transport cargo toward plus end of microtubules, a single dynein moves in opposite direction. How transports wide variety remains an open question. The FTS-Hook-FHIP (“FHF”) complex links mammals fungi. As human cells have three Hooks four FHIP proteins, we hypothesized that combinatorial...

Abstract Pancreatic cancer is the fourth leading cause of mortality in U.S. with only a 25% one-year survival rate and 100% rate. No favorable therapeutic strategy has been established recurrent pancreatic cancer. The cell surface protein CD147 glycoprotein plays central role chemoresistance its upregulation indicates poor prognosis. However, mechanism by which this involved still poorly understood. Here we show that knockdown cells decelerates tumor growth sensitizes to chemotherapeutic...

The Extracellular Matrix Metalloproteinase Inducer (EMMPRIN, CD147) is highly expressed in multiple cancers, particularly pancreatic cancer where it stimulates the secretion of entire classes proteins including matrix matelloproteinases (MMPs) and pro‐inflammatory cytokines. Although, EMMPRIN was initially identified as a cellular transmembrane glycoprotein, extracellular has recently emerged key player disease progression. described study here aims to directly characterize glycosylated...

Abstract Cytoplasmic dynein-1 transports many intracellular cargos towards microtubule minus ends. Dynein is autoinhibited and undergoes conformational changes to form an active complex, consisting of one or two dynein dimers, the dynactin complex activating adaptor(s) 1,2 . The Lissencephaly 1 gene, LIS1 , genetically linked pathway from fungi mammals mutated in patients with neurodevelopmental disease lissencephaly 3–5 Lis1 required for complexes 6–10 but how it does so unclear. Here, we...