Ulk1(S555) inhibition alters nutrient stress response by prioritizing amino acid metabolism

Willoughby, O. S.; Nichenko, A. S.; Brisendine, M. H.; Amiri, N.; Henry, S. N.; Braxton, D. S.; Brown, J. R.; Specht, K. S.; Addington, A. K.; Zaitsev, A. V.; Burrows, S. T.; McMillan, R. P.; Zhang, H.; Tye, S. A.; Najt, C. P.; Craige, S. E.; Rhoads, T. W.; Warren, J. S.; Drake, J. C.

doi:10.1101/2025.06.30.662412

bioRxiv · 10.1101/2025.06.30.662412

Ulk1(S555) inhibition alters nutrient stress response by prioritizing amino acid metabolism

Willoughby, O. S.·Nichenko, A. S.·Brisendine, M. H.·Amiri, N.·Henry, S. N.·Braxton, D. S.·Brown, J. R.·Specht, K. S.·Addington, A. K.·Zaitsev, A. V.·Burrows, S. T.·McMillan, R. P.·Zhang, H.·Tye, S. A.·Najt, C. P.·Craige, S. E.·Rhoads, T. W.·Warren, J. S.·Drake, J. C.

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Abstract

Metabolic flexibility, the capacity to adapt fuel utilization in response to nutrient availability, is essential for maintaining energy homeostasis and preventing metabolic disease. Here, we investigate the role of Ulk1 phosphorylation at serine 555 (S555), a site regulated by AMPK, in coordinating metabolic switching following short-term caloric restriction and fasting. Using Ulk1(S555A) global knock-in mice, we show loss of S555 phosphorylation impairs glucose oxidation in skeletal muscle and liver during short-term CR, despite improved glucose tolerance. Metabolomic, transcriptomic, and mitochondrial respiration analyses suggest a compensatory reliance on autophagy-derived amino acids in Ulk1(S555A) mice. These findings suggest Ulk1(S555) phosphorylation as a critical regulatory event linking nutrient stress to substrate switching. This work highlights an underappreciated role of Ulk1 in maintaining metabolic flexibility, with implications for metabolic dysfunction. Graphical Abstract O_FIG O_LINKSMALLFIG WIDTH=200 HEIGHT=179 SRC="FIGDIR/small/662412v3_ufig1.gif" ALT="Figure 1"> View larger version (22K): org.highwire.dtl.DTLVardef@33a57aorg.highwire.dtl.DTLVardef@b518ccorg.highwire.dtl.DTLVardef@16f2693org.highwire.dtl.DTLVardef@4daac4_HPS_FORMAT_FIGEXP M_FIG C_FIG

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Willoughby, O. S., Nichenko, A. S., Brisendine, M. H., Amiri, N., Henry, S. N., Braxton, D. S., Brown, J. R., Specht, K. S., Addington, A. K., Zaitsev, A. V., Burrows, S. T., McMillan, R. P., Zhang, H., Tye, S. A., Najt, C. P., Craige, S. E., Rhoads, T. W., Warren, J. S., Drake, J. C.. 2025-07-04. Ulk1(S555) inhibition alters nutrient stress response by prioritizing amino acid metabolism. https://doi.org/10.1101/2025.06.30.662412

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