Hrr25 triggers selective autophagy–related pathways by phosphorylating receptor proteins

Tanaka, Chikara; Tan, Li-Jing; Mochida, Keisuke; Kirisako, Hiromi; Koizumi, Michiko; Asai, Eri; Sakoh-Nakatogawa, Machiko; Ohsumi, Yoshinori; Nakatogawa, Hitoshi

doi:10.1083/jcb.201402128

Cited by 109 publications

(118 citation statements)

References 54 publications

Supporting

Mentioning

113

Contrasting

Unclassified

Order By: Relevance

“…The middle band, which was insensitive to Hrr25 depletion, may indicate Atg34 phosphorylation by another kinase. Alternatively, the band might reflect incomplete depletion of Hrr25; consistent with this idea, hrr25-aid cells continued to grow at a slow rate even in the presence of IAA [17]. We also performed in vitro phosphorylation reactions using recombinant Atg34 and Hrr25, which were expressed in and purified from E. coli cells.…”

Section: Hrr25 Phosphorylates Atg34 Under Nitrogen-starvation Conditionsmentioning

confidence: 82%

“…Escherichia coli BL21 cells harboring pGEX-6P-Atg34 (donated by Dr. Nobuo Noda), -Hrr25, or Hrr25 K38A [17], or the pGEX-6P-1 (empty) vector, were cultured at 37°C in LB media (10 mg/ ml tryptone, 5 mg/ml yeast extract, 10 mg/ml NaCl, and 1 mM NaOH) containing 50 lg/ml ampicillin. To induce protein expression, isopropyl b-D-thiogalactopyranoside was added to a final concentration of 0.1 mM.…”

Section: Protein Purificationmentioning

confidence: 99%

See 1 more Smart Citation

Hrr25 phosphorylates the autophagic receptor Atg34 to promote vacuolar transport of α‐mannosidase under nitrogen starvation conditions

2014

Self Cite

View full text Add to dashboard Cite

a b s t r a c tIn Saccharomyces cerevisiae, under nitrogen-starvation conditions, the a-mannosidase Ams1 is recognized by the autophagic receptor Atg34 and transported into the vacuole, where it functions as an active enzyme. In this study, we identified Hrr25 as the kinase that phosphorylates Atg34 under these conditions. Hrr25-mediated phosphorylation does not affect the interaction of Atg34 with Ams1, but instead promotes Atg34 binding to the adaptor protein Atg11, which recruits the autophagy machinery to the Ams1-Atg34 complex, resulting in activation of the vacuolar transport of Ams1. Our findings reveal the regulatory mechanism of a biosynthetic pathway mediated by the autophagy machinery. Structured summary of protein interactions:Hrr25 phosphorylates Atg34 by protein kinase assay (View interaction) Ams1 and Atg34 colocalize by fluorescence microscopy (View interaction) Atg34 physically interacts with Ams1 by anti tag coimmunoprecipitation (View interaction)

show abstract

Section: Hrr25 Phosphorylates Atg34 Under Nitrogen-starvation Conditionsmentioning

confidence: 82%

Section: Protein Purificationmentioning

confidence: 99%

Hrr25 phosphorylates the autophagic receptor Atg34 to promote vacuolar transport of α‐mannosidase under nitrogen starvation conditions

2014

Self Cite

View full text Add to dashboard Cite

show abstract

“…Interestingly, Hrr25 is also involved in many other cellular processes, including autophagy (Tanaka et al, 2014), trafficking through the Golgi (Lord et al, 2011), DNA damage repair (Hoekstra et al, 1991) and mitotic spindle function (Peng et al, 2015b;Petronczki et al, 2006). Whether Hrr25 coordinates any of these activities with CME needs to be determined.…”

Section: Phosphoregulationmentioning

confidence: 99%

Clathrin-mediated endocytosis in budding yeast at a glance

Drubin

Sun

2016

Journal of Cell Science

113

105

View full text Add to dashboard Cite

Clathrin-mediated endocytosis is an essential cellular process that involves the concerted assembly and disassembly of many different proteins at the plasma membrane. In yeast, live-cell imaging has shown that the spatiotemporal dynamics of these proteins is highly stereotypical. Recent work has focused on determining how the timing and functions of endocytic proteins are regulated. In this Cell Science at a Glance article and accompanying poster, we review our current knowledge of the timeline of endocytic site maturation and discuss recent works focusing on how phosphorylation, ubiquitylation and lipids regulate various aspects of the process.

show abstract

“…Several of the Atg proteins are phosphorylated, but until recently it was not known if any of these proteins were Atg1 substrates. Many other protein kinases (i.e., proteins that are not part of the autophagy core machinery) also modulate autophagy activity, [25][26][27][28] but in most cases the relevant targets have not been identified. Atg9 plays a key role in autophagy, and we recently demonstrated that the amount of the Atg9 protein correlates with the rate of autophagosome formation.…”

Section: Determination Of Atg9 Phosphorylation Sitesmentioning

confidence: 99%

Phosphorylation of Atg9 regulates movement to the phagophore assembly site and the rate of autophagosome formation

Feng¹,

Backues²,

Baba³

et al. 2016

Autophagy

View full text Add to dashboard Cite

Macroautophagy is primarily a degradative process that cells use to break down their own components to recycle macromolecules and provide energy under stress conditions, and defects in macroautophagy lead to a wide range of diseases. Atg9, conserved from yeast to mammals, is the only identified transmembrane protein in the yeast core macroautophagy machinery required for formation of the sequestering compartment termed the autophagosome. This protein undergoes dynamic movement between the phagophore assembly site (PAS), where the autophagosome precursor is nucleated, and peripheral sites that may provide donor membrane for expansion of the phagophore. Atg9 is a phosphoprotein that is regulated by the Atg1 kinase. We used stable isotope labeling by amino acids in cell culture (SILAC) to identify phosphorylation sites on this protein and identified an Atg1-independent phosphorylation site at serine 122. A nonphosphorylatable Atg9 mutant showed decreased autophagy activity, whereas the phosphomimetic mutant enhanced activity. Electron microscopy analysis suggests that the different levels of autophagy activity reflect differences in autophagosome formation, correlating with the delivery of Atg9 to the PAS. Finally, this phosphorylation regulates Atg9 interaction with Atg23 and Atg27.

show abstract

Hrr25 triggers selective autophagy–related pathways by phosphorylating receptor proteins

Abstract: The budding yeast kinase Hrr25 regulates two selective autophagy–related pathways by phosphorylating degradation target receptors and thereby promoting their interaction with Atg11 and the formation of autophagosomal membrane.

Cited by 109 publications

References 54 publications

Hrr25 phosphorylates the autophagic receptor Atg34 to promote vacuolar transport of α‐mannosidase under nitrogen starvation conditions

Hrr25 phosphorylates the autophagic receptor Atg34 to promote vacuolar transport of α‐mannosidase under nitrogen starvation conditions

Clathrin-mediated endocytosis in budding yeast at a glance

Phosphorylation of Atg9 regulates movement to the phagophore assembly site and the rate of autophagosome formation

Contact Info

Product

Resources

About