Unc-51 like kinase 1 (ULK1) in silico analysis for biomarker identification: A vital component of autophagy

Randhawa, Rohit; Sehgal, Manika; Singh, Tiratha Raj; Duseja, Ajay; Changotra, Harish

doi:10.1016/j.gene.2015.02.056

Cited by 21 publications

(11 citation statements)

References 49 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…Although it has been reported that ULK3, a member of ULK1 kinase family, could regulate cytokinesis and ATG13 could regulate Colchicinamide-induced mitotic catastrophe [57,61], the roles of autophagy kinase complex ULK1-ATG13 in mitotic regulation are still unclear. While previous reports implicated possible links between ULK1 and CDK1 [62,63], our study here is the first report that demonstrates CDK1 is the upstream kinase for ULK1-ATG13 complex. ULK1-ATG13 phosphorylation could positively regulate autophagy, which promoted Taxol-induced cell death and mitotic slippage.…”

Section: Kinases Involved In Both Autophagy and Mitosiscontrasting

confidence: 48%

CDK1 phosphorylates ULK1-ATG13 complex to regulate mitotic autophagy and Taxol chemosensitivity

Tian

et al. 2019

Preprint

View full text Add to dashboard Cite

ULK1-ATG13 is the most upstream autophagy initiation complex that is phosphorylated by mTORC1 and AMPK to induce autophagy in asynchronous conditions. However, the phospho-regulation and function of ULK1-ATG13 in mitosis and cell cycle remains unknown. Here we show that ULK1-ATG13 complex is differentially regulated throughout the cell cycle. Notably, in mitosis, both ULK1 and ATG13 are highly phosphorylated by CDK1/cyclin B, the key cell cycle machinery. Combining mass spectrometry and site-directed mutagenesis, we found that CDK1-induced ULK1-ATG13 phosphorylation positively regulates mitotic autophagy and Taxol chemosensitivity, and some phosphorylation sites occur in cancer patients. Moreover, double knockout of ULK1 and ATG13 could block cell cycle progression and significantly decrease cancer cell proliferation in cell line and mouse models. Our results not only bridge the mutual regulation between the core machineries of autophagy and mitosis, illustrate the mitotic autophagy regulation mechanism, but also provide ULK1-ATG13 as potential targets for cancer therapy. Immunoprecipitation and Western blotsThe procedure was instructed as previously [68]. Most immunoprecipitation experiments were conducted in HEK-293T derived cell lines due to higher expression level for the exogenous protein. Briefly, HEK-293T cells stably expressing GFP-3FLAG or mULK1-3FLAG were lysed with M-PER supplemented with protease inhibitors and phosphatase inhibitors and centrifuged at 4°C 14,000g for 10 min. The supernatant mixed with pre-incubated Protein G Dynabeads and FLAG antibody at 4°C for 12 h and washed three times with lysis buffer. Then the immunoprecipitate was denatured in 1×SDS-PAGE buffer at 95°C for 7 min and subjected to SDS-PAGE and immunoblotting or Coomassie brilliant blue staining. Apoptosis by flow cytometryCells treated with Taxol combine with or without chloroquine were subjected to apoptosis assay according to the manufacturer instructions (BD Biosciences). Briefly, cells were trypsinized and washed twice with ice cold PBS, and stained with PI and/or FITC-Annexin V in apoptosis buffer for 30 min. The FITC and PE channel were used for flow cytometry detection and the data were analyzed by FlowJo. Cell cycle synchronizationVarious cell synchronization methods are used in this paper, which have been used previously. Briefly, a double-thymidine (2.5 μM) block arrested cells in G1/S border and cells progress through S, G2 and M phase after release. A double-thymidine or single-thymidine block in combination with nocodazole (100 ng/mL) or STLC (5 μM) treatment arrested cells in prometaphase or prophase. A double-thymidine block in combination with RO-3306 (10 μM) treatment arrested cells in late G2 phase and progressed into mitosis after RO-3306 washout for three times with prewarmed PBS. Cell cycle analysis by flow cytometry/FACSCells for cell cycle analysis were trypsinized with 0.25% Trypsin/EDTA. After washing with ice-cold PBS twice, cells were fixed with -20°C 75% ethanol overnight and then stained with PI/RN...

show abstract

Section: Kinases Involved In Both Autophagy and Mitosiscontrasting

confidence: 48%

CDK1 phosphorylates ULK1-ATG13 complex to regulate mitotic autophagy and Taxol chemosensitivity

Tian

et al. 2019

Preprint

View full text Add to dashboard Cite

show abstract

“…Randhawa and colleagues have recently published in silico data that likewise suggests an important role for NF-E2 in ULK1 regulation, albeit this paper does not contain functional data. 34 Ours is the first report demonstrating a role for NF-E2 in erythroid maturation by affecting autophagy genes. Because NIX and ULK1 interact with partner proteins in the autophagy process, 35 their increased expression may disrupt complex formation.…”

Section: Discussionmentioning

confidence: 86%

A novel role for nuclear factor-erythroid 2 in erythroid maturation by modulation of mitochondrial autophagy

et al. 2016

View full text Add to dashboard Cite

show abstract

“…The autophagy process is divided into several stages: (1) Induction, (2) vesicle nucleation, (3) vesicle elongation, and (4) fusion and degradation. Autophagy-related proteins catalyze specific reactions at each step and are essential for the production of autophagic fluxes [ 20 , 62 , 63 , 64 ].…”

Section: Neuronal Death In Nddsmentioning

confidence: 99%

Melatonin and Autophagy in Aging-Related Neurodegenerative Diseases

Luo

Sandhu

Rungratanawanich

et al. 2020

IJMS

107

View full text Add to dashboard Cite

With aging, the nervous system gradually undergoes degeneration. Increased oxidative stress, endoplasmic reticulum stress, mitochondrial dysfunction, and cell death are considered to be common pathophysiological mechanisms of various neurodegenerative diseases (NDDs) such as Alzheimer’s disease (AD), Parkinson’s disease (PD), Huntington’s disease (HD), organophosphate-induced delayed neuropathy (OPIDN), and amyotrophic lateral sclerosis (ALS). Autophagy is a cellular basic metabolic process that degrades the aggregated or misfolded proteins and abnormal organelles in cells. The abnormal regulation of neuronal autophagy is accompanied by the accumulation and deposition of irregular proteins, leading to changes in neuron homeostasis and neurodegeneration. Autophagy exhibits both a protective mechanism and a damage pathway related to programmed cell death. Because of its “double-edged sword”, autophagy plays an important role in neurological damage and NDDs including AD, PD, HD, OPIDN, and ALS. Melatonin is a neuroendocrine hormone mainly synthesized in the pineal gland and exhibits a wide range of biological functions, such as sleep control, regulating circadian rhythm, immune enhancement, metabolism regulation, antioxidant, anti-aging, and anti-tumor effects. It can prevent cell death, reduce inflammation, block calcium channels, etc. In this review, we briefly discuss the neuroprotective role of melatonin against various NDDs via regulating autophagy, which could be a new field for future translational research and clinical studies to discover preventive or therapeutic agents for many NDDs.

show abstract

Unc-51 like kinase 1 (ULK1) in silico analysis for biomarker identification: A vital component of autophagy

Cited by 21 publications

References 49 publications

CDK1 phosphorylates ULK1-ATG13 complex to regulate mitotic autophagy and Taxol chemosensitivity

CDK1 phosphorylates ULK1-ATG13 complex to regulate mitotic autophagy and Taxol chemosensitivity

A novel role for nuclear factor-erythroid 2 in erythroid maturation by modulation of mitochondrial autophagy

Melatonin and Autophagy in Aging-Related Neurodegenerative Diseases

Contact Info

Product

Resources

About