Act88F pseudo-acetylation of lysine 50 (K50) negatively regulates muscle function in Drosophila

Non-histone post-translational modifications (PTMs) are an increasingly important area of study that contribute to our understanding for protein function in the control of health and disease. Protein PTMs, such as acetylation, are emerging as key regulators of striated muscle function, yet our understanding of how non-histone protein acetylation affects muscle physiology remains fragmentary. Previous reports from our lab identified lysine (K) 52 of skeletal muscle alpha actin (ACTA1) to be acetylated, yet no reports have shown a role for ACTA1 acetylation in muscle function. Act88F is 93% homologous to ACTA1 and K50 is identical to K52 of ACTA1. Thus, for these studies, we mutated K50 with a glutamine (Q) on the Act88F gene in the indirect flight muscle (IFM) of Drosophila to mimic acetylation. We examined physiological function (i.e. flight and climbing) as well as biophysical changes between acetylated Act88F and the associated myofibrillar proteins involved in contraction and relaxation (myosin, tropomyosin, and troponin). We report that Act88F K50Q pseudo-acetylation resulted in a flightless phenotype and decreased climbing ability. In addition, we report that Act88F K50Q flies had increased IFM damage as examined via fluorescent imaging. Interestingly, Act88F K50Q did not change actin-myosin binding, actin sliding velocity, or Ca2+ sensitivity in actin-motility assays. However, Act88F pseudo-acetylation did decrease actin filament length, but only after interacting with myosin, implying that Act88F acetylation increases actin filament breaking. These data suggest that Act88F acetylation at K50 worsens muscle function by destabilizing filamentous actin, resulting in muscle tearing and damage to the IFM, which results in loss of flight. This work suggests that actin acetylation, at least on lysine 50, in response to environmental stressors or diet could greatly change muscle function and sarcomere integrity. This research was funded by the Dennis Meiss & Janet Ralston Fund for Nutri-epigenetic Research, the National Institute for General Medical Sciences (NIGMS) of the NIH (P20 GM130459), the National Heart, Lung, and Blood Institute of the NIH (R15 HL143496), National Institute of Aging of the NIH (R21 AG077248) and NSF EPSCOR Track II (OIA-1826801) to B.S.F. This is the full abstract presented at the American Physiology Summit 2024 meeting and is only available in HTML format. There are no additional versions or additional content available for this abstract. Physiology was not involved in the peer review process.