Effects of myosin variants on interacting-heads motif explain distinct hypertrophic and dilated cardiomyopathy phenotypes.

Lorenzo Alamo,James S Ware,Raúl Padrón,Antonio Pinto,Richard E Gillilan,Christine E Seidman,Jonathan G Seidman

doi:10.7554/elife.24634

Abstract

Cardiac β-myosin variants cause hypertrophic (HCM) or dilated (DCM) cardiomyopathy by disrupting sarcomere contraction and relaxation. The locations of variants on isolated myosin head structures predict contractility effects but not the prominent relaxation and energetic deficits that characterize HCM. During relaxation, pairs of myosins form interacting-heads motif (IHM) structures that with other sarcomere proteins establish an energy-saving, super-relaxed (SRX) state. Using a human β-cardiac myosin IHM quasi-atomic model, we defined interactions sites between adjacent myosin heads and associated protein partners, and then analyzed rare variants from 6112 HCM and 1315 DCM patients and 33,370 ExAC controls. HCM variants, 72% that changed electrostatic charges, disproportionately altered IHM interaction residues (expected 23%; HCM 54%, p=2.6×10-19; DCM 26%, p=0.66; controls 20%, p=0.23). HCM variant locations predict impaired IHM formation and stability, and attenuation of the SRX state - accounting for altered contractility, reduced diastolic relaxation, and increased energy consumption, that fully characterizes HCM pathogenesis.

Highlights

Vertebrate skeletal and cardiac muscles produce force through a chemo-mechanical cycle that contracts and relaxes the sarcomere, the contractile unit in all muscle cells
The predicted SAXS profile of PDB 5TBY is consistent with X-ray solution scattering of squid HMM in Ca2+-free (EGTA) conditions (Figure 2B and Figure 2—figure supplement 1) and the computed scattering profile for the model closely agrees with the measured scattering profile of squid heavy meromyosin (HMM) (Gillilan et al, 2013)
Extending the computations to wider angles (q = 1.0 A -1; see Figure 2B and Figure 2—figure supplement 1), revealed that the tarantula PDB 3JBH and the human models are essentially identical above q = 0.4 A -1 and unlikely to be Figure 2 continued on page

Summary

Introduction

Vertebrate skeletal and cardiac muscles produce force through a chemo-mechanical cycle that contracts and relaxes the sarcomere, the contractile unit in all muscle cells. Sarcomeres are composed of inter-digitating thick filaments that contain myosin, comprising a globular head (subfragment 1; S1) and a-helical tail, and thin filaments that contain actin. Interactions between myosin and actin generate the contractile force underpinning sarcomere contraction. Myosin heads comprise a motor domain (MD) that hydrolyses ATP, a regulatory domain (RD) that includes the essential (ELC) and regulatory (RLC) light chains, and a proximal S2 subfragment. Myosin tails are packed in antiparallel formation and shape the thick filament’s backbone, from which myosin heads protrudes every 14.3 nm as crowns formed by three pairs of heads. In some crowns (Zoghbi et al, 2008) the S2 fragment interacts with myosin binding protein-C (MyBP-C), another thick filament protein that modulates sarcomere activity.

Methods

Results

Conclusion