Nanobodies combined with DNA-PAINT super-resolution reveal a staggered titin nanoarchitecture in flight muscles - Aix-Marseille Université Access content directly
Journal Articles eLife Year : 2023

Nanobodies combined with DNA-PAINT super-resolution reveal a staggered titin nanoarchitecture in flight muscles

Renate Rees
  • Function : Author
Jürgen Schünemann
  • Function : Author
Ralf Jungmann
Dirk Görlich

Abstract

Sarcomeres are the force-producing units of all striated muscles. Their nanoarchitecture critically depends on the large titin protein, which in vertebrates spans from the sarcomeric Z-disc to the M-band and hence links actin and myosin filaments stably together. This ensures sarcomeric integrity and determines the length of vertebrate sarcomeres. However, the instructive role of titins for sarcomeric architecture outside of vertebrates is not as well understood. Here, we used a series of nanobodies, the Drosophila titin nanobody toolbox, recognising specific domains of the two Drosophila titin homologs Sallimus and Projectin to determine their precise location in intact flight muscles. By combining nanobodies with DNA-PAINT super-resolution microscopy, we found that, similar to vertebrate titin, Sallimus bridges across the flight muscle I-band, whereas Projectin is located at the beginning of the A-band. Interestingly, the ends of both proteins overlap at the I-band/A-band border, revealing a staggered organisation of the two Drosophila titin homologs. This architecture may help to stably anchor Sallimus at the myosin filament and hence ensure efficient force transduction during flight. Editor's evaluation This landmark study combines two advanced technologies, namely, nanobodies and DNA-PAINT, to define the position of several subdomains of the two fly titin homologs in adult flight muscles. Their results provide compelling evidence that Sallimus can bridge the Z-disk with the beginning of the myosin A-band. Furthermore, their results convincingly establish that Sallimus and Projectin partially overlap at the beginning of the A-band. The work should appeal to people interested in muscle biology and more generally to people interested in providing high-resolution images of long proteins.
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hal-04290002 , version 1 (16-11-2023)

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Florian Schueder, Pierre Mangeol, Eunice Hoyee Chan, Renate Rees, Jürgen Schünemann, et al.. Nanobodies combined with DNA-PAINT super-resolution reveal a staggered titin nanoarchitecture in flight muscles. eLife, 2023, 12, ⟨10.7554/eLife.79344⟩. ⟨hal-04290002⟩
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