[ASAP] Cas14: Big Advances from Small CRISPR Proteins

February 11th, 2019 by David F. Savage

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Biochemistry
DOI: 10.1021/acs.biochem.9b00035

[ASAP] Thrombin Exosite Maturation and Ligand Binding at ABE II Help Stabilize PAR-Binding Competent Conformation at ABE I

February 11th, 2019 by Ramya Billur, T. Michael Sabo, Muriel C. Maurer

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Biochemistry
DOI: 10.1021/acs.biochem.8b00943

[ASAP] Combining 26s rDNA and the Cre-loxP System for Iterative Gene Integration and Efficient Marker Curation in <italic toggle=”yes”>Yarrowia lipolytica</italic>

February 8th, 2019 by Yongkun Lv, Harley Edwards, Jingwen Zhou, Peng Xu

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ACS Synthetic Biology
DOI: 10.1021/acssynbio.8b00535
  • Posted in ACS Synthetic Biology, Publications
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Author Correction: Complete reconstitution of the diverse pathways of gentamicin B biosynthesis

February 6th, 2019 by Yeon Hee Ban

Author Correction: Complete reconstitution of the diverse pathways of gentamicin B biosynthesis

Author Correction: Complete reconstitution of the diverse pathways of gentamicin B biosynthesis, Published online: 06 February 2019; doi:10.1038/s41589-019-0232-7

Author Correction: Complete reconstitution of the diverse pathways of gentamicin B biosynthesis
  • Posted in Nat Chem Biol, Publications
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[ASAP] Shaping the Future of Protein Engineering

February 6th, 2019 by Dominic J. Glover, Dawei Xu, Douglas S. Clark

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Biochemistry
DOI: 10.1021/acs.biochem.8b01322

15-Deoxy-{Delta}12,14-prostaglandin J2 promotes phosphorylation of eukaryotic initiation factor 2{alpha} and activates the integrated stress response [Neurobiology]

February 5th, 2019 by Devin Tauber, Roy Parker

Stress granules (SGs) are cytoplasmic RNA-protein aggregates formed in response to inhibition of translation initiation. SGs contribute to the stress response and are implicated in a variety of diseases including cancer and some forms of neurodegeneration. Neurodegenerative diseases often involve chronic phosphorylation of eukaryotic initiation factor 2α (eIF2α), with deletions of eIF2α kinases or treatment with eiF2α kinase inhibitors being protective in some animal models of disease. However, how and why the integrated stress response (ISR) is activated in different forms of neurodegeneration remains unclear. Since neuroinflammation is common to many neurodegenerative diseases, we hypothesized that inflammatory factors contribute to ISR activation in a cell non-autonomous manner. Using fluorescence microscopy and immunoblotting, we show here that the endogenously produced product of inflammation, 15-Deoxy-Δ12,14-prostaglandin J2 (15-d-PGJ2), triggers eIF2α phosphorylation thereby activating the ISR, repressing bulk translation, and triggering stress granule formation. Our findings define a mechanism by which inflammation activates the ISR in a cell non-autonomous manner and suggest that inhibition of 15-d-PGJ2 production might be a useful therapeutic strategy in some neuroinflammatory contexts.
  • Posted in Journal of Biological Chemistry, Publications
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The FtsK-like motor TraB is a DNA-dependent ATPase that forms higher-order assemblies [Microbiology]

February 5th, 2019 by Eric Amado, Gunther Muth, Ignacio Arechaga, Elena Cabezon

TraB is an FtsK-like DNA translocase responsible for conjugative plasmid transfer in mycelial Streptomyces. Unlike other conjugative systems, which depend on a type IV secretion system, Streptomyces requires only TraB protein to transfer the plasmid as dsDNA. The γ-domain of this protein specifically binds to repeated 8-bp motifs on the plasmid sequence, following a mechanism that is reminiscent of the FtsK/SpoIIIE chromosome segregation system. In this work, we purified and characterized the enzymatic activity of TraB, revealing that it is a DNA-dependent ATPase that is highly stimulated by dsDNA substrates. Interestingly, we found that unlike the SpoIIIE protein, the γ-domain of TraB does not confer sequence-specific ATPase stimulation. We also found that TraB binds G-quadruplex DNA structures with higher affinity than TraB-recognition sequences (TRSs). An EM-based structural analysis revealed that TraB tends to assemble as large complexes comprising four TraB hexamers, which might be a prerequisite for DNA translocation across cell membranes. In summary, our findings shed light on the molecular mechanism used by the DNA-translocating motor TraB, which may be shared by other membrane-associated machineries involved in DNA binding and translocation.

[ASAP] Redirecting Metabolic Flux <italic toggle=”yes”>via</italic> Combinatorial Multiplex CRISPRi-Mediated Repression for Isopentenol Production in <italic toggle=”yes”>Escherichia coli</italic>

February 5th, 2019 by Tian Tian, Jing Wei Kang, Aram Kang, Taek Soon Lee

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ACS Synthetic Biology
DOI: 10.1021/acssynbio.8b00429
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A chemoproteomic portrait of the oncometabolite fumarate

February 4th, 2019 by Rhushikesh A. Kulkarni

A chemoproteomic portrait of the oncometabolite fumarate

A chemoproteomic portrait of the oncometabolite fumarate, Published online: 04 February 2019; doi:10.1038/s41589-018-0217-y

Chemoproteomic mapping of fumarate sensitive cysteines in a hereditary leiomyomatosis and renal cell carcinoma cell line revealed a critical cysteine in the protein–protein interaction interface of the SWI–SNF complex.

Small-molecule allosteric inhibitors of BAX

February 4th, 2019 by Thomas P. Garner

Small-molecule allosteric inhibitors of BAX

Small-molecule allosteric inhibitors of BAX, Published online: 04 February 2019; doi:10.1038/s41589-018-0223-0

Using biochemical and NMR studies, a class of small-molecule inhibitors termed BAX activation inhibitors were found to bind directly to a previously unrecognized pocket of inactive BAX and allosterically inhibit conformational changes in BAX.