Isoform-specific functions of synaptopodin-2 variants in cytoskeleton stabilization and autophagy regulation in muscle under mechanical stress

Exp Cell Res. 2021 Nov 15;408(2):112865. doi: 10.1016/j.yexcr.2021.112865. Epub 2021 Oct 11.

Abstract

Protein homeostasis (proteostasis) in multicellular organisms depends on the maintenance of force-bearing and force-generating cellular structures. Within myofibrillar Z-discs of striated muscle, isoforms of synaptopodin-2 (SYNPO2/myopodin) act as adapter proteins that are engaged in proteostasis of the actin-crosslinking protein filamin C (FLNc) under mechanical stress. SYNPO2 directly binds F-actin, FLNc and α-actinin and thus contributes to the architectural features of the actin cytoskeleton. By its association with autophagy mediating proteins, i.e. BAG3 and VPS18, SYNPO2 is also engaged in protein quality control and helps to target mechanical unfolded and damaged FLNc for degradation. Here we show that deficiency of all SYNPO2-isoforms in myotubes leads to decreased myofibrillar stability and deregulated autophagy under mechanical stress. In addition, isoform-specific proteostasis functions were revealed. The PDZ-domain containing variant SYNPO2b and the shorter, PDZ-less isoform SYNPO2e both localize to Z-discs. Yet, SYNPO2e is less stably associated with the Z-disc than SYNPO2b, and is dynamically transferred into FLNc-containing myofibrillar lesions under mechanical stress. SYNPO2e also recruits BAG3 into these lesions via interaction with the WW domain of BAG3. Our data provide evidence for a role of myofibrillar lesions as a transient quality control compartment essential to prevent and repair contraction-induced myofibril damage in muscle and indicate an important coordinating activity for SYNPO2 therein.

Keywords: Autophagy; Electrical pacing stimulation; Exercise; Myofibril remodeling; Myopodin/synaptopodin-2 isoforms; Sarcomeric Z-disc.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Actin Cytoskeleton / genetics
  • Actinin / genetics
  • Actins / genetics
  • Adaptor Proteins, Signal Transducing / genetics*
  • Animals
  • Apoptosis Regulatory Proteins / genetics*
  • Autophagy / genetics
  • Cell Line
  • Cytoskeleton / genetics
  • Humans
  • Mice
  • Microfilament Proteins / genetics*
  • Muscle Fibers, Skeletal / metabolism
  • Muscle, Skeletal / metabolism*
  • Muscle, Striated / metabolism
  • Myofibrils / genetics
  • Myofibrils / metabolism
  • PDZ Domains / genetics
  • Protein Isoforms / genetics
  • Stress, Mechanical*
  • Synaptophysin / genetics
  • Vesicular Transport Proteins / genetics*

Substances

  • Actins
  • Adaptor Proteins, Signal Transducing
  • Apoptosis Regulatory Proteins
  • Bag3 protein, mouse
  • Microfilament Proteins
  • Protein Isoforms
  • Synaptophysin
  • Synpo2 protein, mouse
  • Synpr protein, mouse
  • Vesicular Transport Proteins
  • Vps18 protein, mouse
  • Actinin