Human deafness mutation E385D disrupts the mechanochemical coupling and subcellular targeting of myosin-1a.

Abstract:

:Missense mutations in the membrane-binding actin-based motor protein, myosin-1a (Myo1a), have recently been linked to sensorineural deafness in humans. One of these mutations, E385D, impacts a residue in the switch II region of the motor domain that is present in virtually all members of the myosin superfamily. We sought to examine the impact of E385D on the function of Myo1a, both in terms of mechanochemical activity and ability to target to actin-rich microvilli in polarized epithelial cells. While E385D-Myo1a demonstrated actin-activated ATPase activity, the V(MAX) was reduced threefold relative to wild-type. Despite maintaining an active mechanochemical cycle, E385D-Myo1a was unable to move actin in the sliding filament assay. Intriguingly, when an enhanced-green-fluorescent-protein-tagged form of E385D-Myo1a was stably expressed in polarized epithelial cells, this mutation abolished the microvillar targeting normally demonstrated by wild-type Myo1a. Notably, these data are the first to suggest that mechanical activity is essential for proper localization of Myo1a in microvilli. These studies also provide a unique example of how even the most mild substitution of invariant switch II residues can effectively uncouple enzymatic and mechanical activity of the myosin motor domain.

journal_name

Biophys J

journal_title

Biophysical journal

authors

Yengo CM,Ananthanarayanan SK,Brosey CA,Mao S,Tyska MJ

doi

10.1529/biophysj.107.122689

subject

Has Abstract

pub_date

2008-01-15 00:00:00

pages

L5-7

issue

2

eissn

0006-3495

issn

1542-0086

pii

S0006-3495(08)70708-2

journal_volume

94

pub_type

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