An actin-dependent annexin complex mediates plasma membrane repair in muscle.
Disruption of the plasma membrane often accompanies cellular injury, and in muscle, plasma membrane resealing is essential for efficient recovery from injury. Muscle contraction, especially of lengthened muscle, disrupts the sarcolemma. To define the molecular machinery that directs repair, we applied laser wounding to live mammalian myofibers and assessed translocation of fluorescently tagged proteins using high-resolution microscopy. Within seconds of membrane disruption, annexins A1, A2, A5, and A6 formed a tight repair "cap." Actin was recruited to the site of damage, and annexin A6 cap formation was both actin dependent and Ca(2+) regulated. Repair proteins, including dysferlin, EHD1, EHD2, MG53, and BIN1, localized adjacent to the repair cap in a "shoulder" region enriched with phosphatidlyserine. Dye influx into muscle fibers lacking both dysferlin and the related protein myoferlin was substantially greater than control or individual null muscle fibers, underscoring the importance of shoulder-localized proteins. These data define the cap and shoulder as subdomains within the repair complex accumulating distinct and nonoverlapping components.
Center for Genetic Medicine, Northwestern University, Chicago, IL 60611.
Alexis R
AR
Demonbreun
Center for Genetic Medicine, Northwestern University, Chicago, IL 60611.
Mattia
M
Quattrocelli
Center for Genetic Medicine, Northwestern University, Chicago, IL 60611.
David Y
DY
Barefield
Center for Genetic Medicine, Northwestern University, Chicago, IL 60611.
Madison V
MV
Allen
Center for Genetic Medicine, Northwestern University, Chicago, IL 60611 Department of Pathology, The University of Chicago, Chicago, IL 60637.
Kaitlin E
KE
Swanson
Center for Genetic Medicine, Northwestern University, Chicago, IL 60611 elizabeth.mcnally@northwestern.edu alexis.demonbreun@northwestern.edu.
Elizabeth M
EM
McNally
D000199:Actins / Q000378:metabolism*; D000818:Animals; D017302:Annexins / Q000378:metabolism*; D002118:Calcium / Q000378:metabolism; D002462:Cell Membrane / Q000378:metabolism* / Q000502:physiology*; D051379:Mice; D018485:Muscle Fibers, Skeletal / Q000378:metabolism* / Q000502:physiology*; D021381:Protein Transport / Q000502:physiology; D012508:Sarcolemma / Q000378:metabolism; D014945:Wound Healing / Q000502:physiology
D000199:Actins; D017302:Annexins; D002118:Calcium