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- W2026092029 abstract "Class III myosins (myo3A and myo3B) contain a N-terminal kinase domain, central motor domain, and an isoform specific C-terminal tail. Myo3A contains a longer tail and has an actin-binding motif at the C-terminus that is not present in the shorter tail of myo3B. Modifications to the myo3A gene result in late onset human deafness (DFNB30), suggesting myo3B may be able to compensate for myo3A only early in life. Thus, it is critical to understand differences in the motor properties of myo3A and myo3B. We investigated the in vitro motility of myo3A and myo3B, each with the kinase domain removed and containing two IQ domains and a C-terminal GFP fusion (myo3AΔK.GFP and myo3BΔK.GFP). The sliding velocity measured in the in vitro motility assay was ∼20% faster in myo3AΔK.GFP (76 nm/sec) compared to myo3BΔK.GFP (62 nm/sec). The sliding velocity correlated well with the maximum actin-activated ATPase activity (kcat = 1.0 sec−1 and 0.7 sec−1, respectively). Interestingly, myo3B.ΔK.GFP contained a 25-fold weaker actin affinity compared to myo3A.ΔK.GFP as assessed by the actin-dependence of the ATPase activity (KATPase = 3 and 76 µM, respectively). We transfected N-terminally GFP tagged myo3A.ΔK containing the full length tail domain into COS7 cells and examined the efficiency of localizing to the filopodia tips by examining the tip to cell body ratio. Since the tail actin binding motif is required for localization to filopodia tips, we generated a chimera that contained the myo3B motor and the actin binding motif of the myo3A tail. The tip localization efficiency was approximately 17% (p<0.05) reduced in myo3B compared to myo3A. Thus myo3B is a slightly slower motor with a weaker actin affinity compared to myo3A, which may explain why it is unable to compensate for the loss of myo3A in DFNB30." @default.
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- W2026092029 date "2014-01-01" @default.
- W2026092029 modified "2023-09-30" @default.
- W2026092029 title "Class III Myosin Motor Activity Correlates with Localization in Actin Protrusions" @default.
- W2026092029 doi "https://doi.org/10.1016/j.bpj.2013.11.1018" @default.
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