APOL1–Mediated Cell Injury Involves Disruption of Conserved Trafficking Processes

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An optimized transgenesis system for Drosophila using germ-line-specific phiC31 integrases.

Johannes Bischof, Robert Maeda, Monika Hediger … (2007)

Germ-line transformation via transposable elements is a powerful tool to study gene function in Drosophila melanogaster. However, some inherent characteristics of transposon-mediated transgenesis limit its use for transgene analysis. Here, we circumvent these limitations by optimizing a phiC31-based integration system. We generated a collection of lines with precisely mapped attP sites that allow the insertion of transgenes into many different predetermined intergenic locations throughout the fly genome. By using regulatory elements of the nanos and vasa genes, we established endogenous sources of the phiC31 integrase, eliminating the difficulties of coinjecting integrase mRNA and raising the transformation efficiency. Moreover, to discriminate between specific and rare nonspecific integration events, a white gene-based reconstitution system was generated that enables visual selection for precise attP targeting. Finally, we demonstrate that our chromosomal attP sites can be modified in situ, extending their scope while retaining their properties as landing sites. The efficiency, ease-of-use, and versatility obtained here with the phiC31-based integration system represents an important advance in transgenesis and opens up the possibility of systematic, high-throughput screening of large cDNA sets and regulatory elements.

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Dynamics and diversity in autophagy mechanisms: lessons from yeast.

Hitoshi Nakatogawa, Kuninori Suzuki, Yoshiaki Kamada … (2009)

Autophagy is a fundamental function of eukaryotic cells and is well conserved from yeast to humans. The most remarkable feature of autophagy is the synthesis of double membrane-bound compartments that sequester materials to be degraded in lytic compartments, a process that seems to be mechanistically distinct from conventional membrane traffic. The discovery of autophagy in yeast and the genetic tractability of this organism have allowed us to identify genes that are responsible for this process, which has led to the explosive growth of this research field seen today. Analyses of autophagy-related (Atg) proteins have unveiled dynamic and diverse aspects of mechanisms that underlie membrane formation during autophagy.

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Differential regulation of distinct Vps34 complexes by AMPK in nutrient stress and autophagy.

Joungmok Kim, Young Chul Kim, Chong Fang … (2013)

Autophagy is a stress response protecting cells from unfavorable conditions, such as nutrient starvation. The class III phosphatidylinositol-3 kinase, Vps34, forms multiple complexes and regulates both intracellular vesicle trafficking and autophagy induction. Here, we show that AMPK plays a key role in regulating different Vps34 complexes. AMPK inhibits the nonautophagy Vps34 complex by phosphorylating T163/S165 in Vps34 and therefore suppresses overall PI(3)P production and protects cells from starvation. In parallel, AMPK activates the proautophagy Vps34 complex by phosphorylating S91/S94 in Beclin1 to induce autophagy. Atg14L, an autophagy-essential gene present only in the proautophagy Vps34 complex, inhibits Vps34 phosphorylation but increases Beclin1 phosphorylation by AMPK. As such, Atg14L dictates the differential regulation (either inhibition or activation) of different Vps34 complexes in response to glucose starvation. Our study reveals an intricate molecular regulation of Vps34 complexes by AMPK in nutrient stress response and autophagy. Copyright © 2013 Elsevier Inc. All rights reserved.

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Author and article information

Journal

Title: Journal of the American Society of Nephrology

Abbreviated Title: JASN

Publisher: American Society of Nephrology (ASN)

ISSN (Print): 1046-6673

ISSN (Electronic): 1533-3450

Publication date Created: March 31 2017

Publication date (Print): April 18 2017

Volume: 28

Issue: 4

Pages: 1117-1130

Article

DOI: 10.1681/ASN.2016050546

PMC ID: 5373454

PubMed ID: 27864431

SO-VID: c009ba92-89bf-487c-9f4a-02f31502d0da

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APOL1–Mediated Cell Injury Involves Disruption of Conserved Trafficking Processes

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Anti-Trafficking Review

Most cited references 65

An optimized transgenesis system for Drosophila using germ-line-specific phiC31 integrases.

Dynamics and diversity in autophagy mechanisms: lessons from yeast.

Differential regulation of distinct Vps34 complexes by AMPK in nutrient stress and autophagy.

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