Autophagy inhibition by biotin elicits endoplasmic reticulum stress to differentially regulate adipocyte lipid and protein synthesis

  • Selvam Senthilraja 
  • Ramaian Santhaseela Anand 
  • Ganesan Dhasarathan 
  • Rajasekaran Sudarshana 
  • Jayavelu Tamilselvan Email author
Original Paper


Biotin is an indispensable adipogenic agent, and its ability to coordinate carbohydrate, lipid, and amino acid metabolism sensitizes insulin signaling in adipocytes. This enables the organism to adapt and survive under nutrient stress by synthesis and storage of lipids. Biotin deficiency mimics insulin resistance with alterations in cellular intermediary metabolism. Though the mechanism of lipogenesis is well established across cell types, considering its predisposition to accumulate only lipids, it is necessary to elucidate the mechanism that minimizes the effects of biotin on adipocyte protein synthesis. In order to determine the differential metabolic phenotype by biotin, the primary cultures of adipocytes were induced to differentiate in the presence and absence of excess biotin. Serum pre-incubated with avidin was used to limit biotin availability in cultured cells. Biotin restricts cellular signaling associated with protein synthesis without altering total protein content. The decline in autophagy elicits endoplasmic reticulum stress to inhibit protein synthesis by eIF2α phosphorylation possibly via accumulation of misfolded/long-lived proteins. Furthermore, the compensatory increase in Unc51 like autophagy activating kinase 1 possibly competes with eukaryotic initiation factor 4E-binding protein 1 and ribosomal p70 S6kinase phosphorylation by mechanistic targets of rapamycin complex 1 to uncouple its effect on protein synthesis. In conclusion, autophagy inhibition by biotin uncouples protein synthesis to promote lipogenesis by eliciting endoplasmic reticulum stress and differential phosphorylation of mechanistic targets of rapamycin complex 1 substrates.


Biotin Autophagy ER stress mTORC1 Amino acids Protein synthesis 



We would also like to acknowledge DBT-BUILDER program (BT/PR12153/INF/22/200/2014) for providing HPLC instrumentation and Dr. S. Meenakshisundaram’s research group for their assistance with HPLC analysis. The authors SRS and ARS acknowledge ICMR, Government of India for Senior Research Fellowship award.

Funding information

This study was supported in part by grants-in-aid for research from DST-SERB (SR/S0/HS/0051/2012 and EMR/2016/003276).

Compliance with ethical standards

The protocols for animal maintenance and usage were approved by the Institutional Animal Ethics Committee.

Supplementary material

12192_2018_967_Fig5_ESM.png (1.4 mb)

(PNG 1417 kb)

12192_2018_967_MOESM1_ESM.tif (24.1 mb)
High Resolution Image (TIF 24684 kb)


  1. Averous J, Lambert-Langlais S, Mesclon F, Carraro V, Parry L, Jousse C, Bruhat A, Maurin AC, Pierre P, Proud CG, Fafournoux P (2016) GCN2 contributes to mTORC1 inhibition by leucine deprivation through an ATF4 independent mechanism. Sci Rep 6:1–10. CrossRefGoogle Scholar
  2. Blanchette-Mackie EJ, Dwyer NK, Barber T, Coxey RA, Takeda T, Rondinone CM, Theodorakis JL, Greenberg AS, Londos C (1995) Perilipin is located on the surface layer of intracellular lipid droplets in adipocytes. J Lipid Res 36:1211–1226Google Scholar
  3. Boeckx RL, Dakshinamurti K (1974) Biotin-mediated protein biosynthesis. Biochem J 140:549–556CrossRefGoogle Scholar
  4. Chandler CS, Ballard FJ (1985) Distribution and degradation of biotin-containing carboxylases in human cell lines. Biochem J 232:385–393. CrossRefGoogle Scholar
  5. Chen GL, Sutrina SL, Frayer KL, Chen WW (1986) Effects of lysosomotropic agents on lipogenesis. Arch Biochem Biophys 245:66–75. CrossRefGoogle Scholar
  6. Cheng T, Sudderth J, Yang C, Mullen AR, Jin ES, Mates JM, DeBerardinis RJ (2011) Pyruvate carboxylase is required for glutamine-independent growth of tumor cells. Proc Natl Acad Sci U S A 108:8674–8679CrossRefGoogle Scholar
  7. Dakshinamurti K, Desjardins PR (1968) Lipogenesis in biotin deficiency. Can J Biochem 46:1261–1267. CrossRefGoogle Scholar
  8. Dakshinamurti K, Modi VV, Mistry SP (1968) Some aspects of carbohydrates metabolism in biotin-deficient rats. Proc Soc Exp Biol Med 127:396–400CrossRefGoogle Scholar
  9. Damiano F, Alemanno S, Gnoni GV, Siculella L (2010) Translational control of the sterol-regulatory transcription factor SREBP-1 mRNA in response to serum starvation or ER stress is mediated by an internal ribosome entry site. Biochem J 429:603–612. CrossRefGoogle Scholar
  10. Damiano F, Rochira A, Tocci R, Alemanno S, Gnoni A, Siculella L (2013) hnRNP A1 mediates the activation of the IRES-dependent SREBP-1a mRNA translation in response to endoplasmic reticulum stress. Biochem J 449:543–553. CrossRefGoogle Scholar
  11. Dey M, Cao C, Sicheri F, Dever TE (2007) Conserved intermolecular salt bridge required for activation of protein kinases PKR, GCN2, and PERK. J Biol Chem 282:6653–6660. CrossRefGoogle Scholar
  12. Dibble CC, Manning BD (2013) Signal integration by mTORC1 coordinates nutrient input with biosynthetic output. Nat Cell Biol 15:555–564. CrossRefGoogle Scholar
  13. Dunlop EA, Hunt DK, Acosta-Jaquez HA, Fingar DC, Tee AR (2011) ULK1 inhibits mTORC1 signaling, promotes multisite raptor phosphorylation and hinders substrate binding. Autophagy 7:737–747. CrossRefGoogle Scholar
  14. Eleftheriadis T, Pissas G, Antoniadi G et al (2016) Differential effects of the two amino acid sensing systems, the GCN2 kinase and the mTOR complex 1, on primary human alloreactive CD4+T-cells. Int J Mol Med 37:1412–1420. CrossRefGoogle Scholar
  15. Fang DL, Wan Y, Shen W, Cao J, Sun ZX, Yu HH, Zhang Q, Cheng WH, Chen J, Ning B (2013) Endoplasmic reticulum stress leads to lipid accumulation through upregulation of SREBP-1c in normal hepatic and hepatoma cells. Mol Cell Biochem 381:127–137. CrossRefGoogle Scholar
  16. Harding HP, Zhang Y, Bertolotti A, Zeng H, Ron D (2000) Perk is essential for translational regulation and cell survival during the unfolded protein response. Mol Cell 5:897–904. CrossRefGoogle Scholar
  17. Harding HP, Zhang Y, Zeng H, Novoa I, Lu PD, Calfon M, Sadri N, Yun C, Popko B, Paules R, Stojdl DF, Bell JC, Hettmann T, Leiden JM, Ron D (2003) An integrated stress response regulates amino acid metabolism and resistance to oxidative stress. Mol Cell 11:619–633. CrossRefGoogle Scholar
  18. Hariharan M, Naga S, VanNoord T (1993) Systematic approach to the development of plasma amino acid analysis by high-performance liquid chromatography with ultraviolet detection with precolumn derivatization using phenyl isothiocyanate. J Chromatogr 621:15–22CrossRefGoogle Scholar
  19. Hymes J, Wolf B (1999) Human biotinidase isn’t just for recycling biotin. J Nutr 129:485S–489SCrossRefGoogle Scholar
  20. Jacobs R, Kilburn E, Majerus PW (1970) Acetyl coenzyme a carboxylase. The effects of biotin deficiency on enzyme in rat liver and adipose tissue. J Biol Chem 245:6462–6467Google Scholar
  21. Kim J, Kundu M, Viollet B, Guan KL (2011) AMPK and mTOR regulate autophagy through direct phosphorylation of Ulk1. Nat Cell Biol 13:132–141. CrossRefGoogle Scholar
  22. Klemm RW, Norton JP, Cole RA, Li CS, Park SH, Crane MM, Li L, Jin D, Boye-Doe A, Liu TY, Shibata Y, Lu H, Rapoport TA, Farese RV Jr, Blackstone C, Guo Y, Mak HY (2013) A conserved role for Atlastin GTPases in regulating lipid droplet size. Cell Rep 3:1465–1475. CrossRefGoogle Scholar
  23. Kuri-Harcuch W, Wise LS, Green H (1978) Interruption of the adipose conversion of 3T3 cells by biotin deficiency: differentiation without triglyceride accumulation. Cell 14:53–59. CrossRefGoogle Scholar
  24. Lowry OH, Rosebrough NJ, Farr AL, Randall RJ (1951) Protein measurement with the Folin phenol reagent. J Biol Chem 193:265–275Google Scholar
  25. Lynen F (1967) The role of biotin-dependent carboxylations in biosynthetic reactions. Biochem J 102:381–400. CrossRefGoogle Scholar
  26. Ma K, Vattem KM, Wek RC (2002) Dimerization and release of molecular chaperone inhibition facilitate activation of eukaryotic initiation factor-2 kinase in response to endoplasmic reticulum stress. J Biol Chem 277:18728–18735. CrossRefGoogle Scholar
  27. Martin S, Parton RG (2006) Lipid droplets: a unified view of dynamic organelle. Mol Cell 7:373–378. Google Scholar
  28. Matsuo H, Li H, McGuire AM et al (1997) Structure of translation factor elF4E bound to m7GDP and interaction with 4E-binding protein. Nat Struct Biol 4:717–724. CrossRefGoogle Scholar
  29. Meijer AJ, Codogno P (2008) Autophagy: a sweet process in diabetes. Cell Metab 8:275–276. CrossRefGoogle Scholar
  30. Negrel R, Dani C (2001) Cultures of adipose precursor cells and cells of clonal lines from animal white adipose tissue. Methods Mol Biol 155:225–237. Google Scholar
  31. Ogata M, Hino S, Saito A, Morikawa K, Kondo S, Kanemoto S, Murakami T, Taniguchi M, Tanii I, Yoshinaga K, Shiosaka S, Hammarback JA, Urano F, Imaizumi K (2006) Autophagy is activated for cell survival after endoplasmic reticulum stress. Mol Cell Biol 26:9220–9231. CrossRefGoogle Scholar
  32. Okayasu T, Ikeda M, Akimoto K, Sorimachi K (1997) The amino acid composition of mammalian and bacterial cells. Amino Acids 13:379–391. CrossRefGoogle Scholar
  33. Onodera J, Ohsumi Y (2005) Autophagy is required for maintenance of amino acid levels and protein synthesis under nitrogen starvation. J Biol Chem 280:31582–31586. CrossRefGoogle Scholar
  34. Orso G, Pendin D, Liu S, Tosetto J, Moss TJ, Faust JE, Micaroni M, Egorova A, Martinuzzi A, McNew JA, Daga A (2009) Homotypic fusion of ER membranes requires the dynamin-like GTPase Atlastin. Nature 460:978–983. CrossRefGoogle Scholar
  35. Sarjeant K, Stephens JM (2012) Adipogenesis. Cold Spring Harb Perspect Biol 4:a008417. CrossRefGoogle Scholar
  36. Schuck S, Prinz WA, Thorn KS, Voss C, Walter P (2009) Membrane expansion alleviates endoplasmic reticulum stress independently of the unfolded protein response. J Cell Biol 187:525–536. CrossRefGoogle Scholar
  37. Tsukiyama-Kohara K, Katsume A, Kimura K, Saito M, Kohara M (2013) 4E-BP1 regulates the differentiation of white adipose tissue. Genes Cells 18:602–607. CrossRefGoogle Scholar
  38. Wek RC, Jiang H-Y, Anthony TG (2006) Coping with stress: eIF2 kinases and translational control. Biochem Soc Trans 34:7. CrossRefGoogle Scholar
  39. Wong PM, Feng Y, Wang J, Shi R, Jiang X (2015) Regulation of autophagy by coordinated action of mTORC1 and protein phosphatase 2A. Nat Commun 6:8048CrossRefGoogle Scholar
  40. Yu L, McPhee CK, Zheng L et al (2010) Termination of autophagy and reformation of lysosomes regulated by mTOR. Nature 465:942–946. CrossRefGoogle Scholar
  41. Yuan W, Guo S, Gao J, Zhong M, Yan G, Wu W, Chao Y, Jiang Y (2017) General control nonderepressible 2 (GCN2) kinase inhibits target of rapamycin complex 1 in response to amino acid starvation in saccharomyces cerevisiae. J Biol Chem 292:2660–2669. CrossRefGoogle Scholar
  42. Zhang HH, Huang J, Düvel K, Boback B, Wu S, Squillace RM, Wu CL, Manning BD (2009) Insulin stimulates adipogenesis through the Akt-TSC2-mTORC1 pathway. PLoS One 4:e6189. CrossRefGoogle Scholar
  43. Zhao J, Brault JJ, Schild A, Cao P, Sandri M, Schiaffino S, Lecker SH, Goldberg AL (2007) FoxO3 coordinately activates protein degradation by the Autophagic/lysosomal and proteasomal pathways in atrophying muscle cells. Cell Metab 6:472–483. CrossRefGoogle Scholar
  44. Zhao J, Zhai B, Gygi SP, Goldberg AL (2015) mTOR inhibition activates overall protein degradation by the ubiquitin proteasome system as well as by autophagy. Proc Natl Acad Sci U S A 112:15790–15797. CrossRefGoogle Scholar
  45. Zhu G, Ye R, Jung DY, Barron E, Friedline RH, Benoit VM, Hinton DR, Kim JK, Lee AS (2013) GRP78 plays an essential role in adipogenesis and postnatal growth in mice. FASEB J 27:955–964. CrossRefGoogle Scholar

Copyright information

© Cell Stress Society International 2019

Authors and Affiliations

  • Selvam Senthilraja 
    • 1
  • Ramaian Santhaseela Anand 
    • 1
  • Ganesan Dhasarathan 
    • 1
  • Rajasekaran Sudarshana 
    • 1
  • Jayavelu Tamilselvan 
    • 1
    Email author
  1. 1.Centre for BiotechnologyAnna UniversityChennaiIndia

Personalised recommendations