期刊名称：Proceedings of the National Academy of Sciences
印刷版ISSN：0027-8424
电子版ISSN：1091-6490
出版年度：2022
卷号：119
期号：8
DOI：10.1073/pnas.2120962119
语种：English
出版社：The National Academy of Sciences of the United States of America
摘要：Significance
It has been suggested for more than 50 y that the fastest myosin in the biological world with a velocity of 70 μm s
−1 exists in the alga
Chara, because cytoplasmic streaming with a velocity of 70 μm s
−1 occurs in
Chara cells. However, a myosin with that velocity has not yet been identified. In this work, we succeeded in cloning a myosin XI with a velocity of 60 μm s
−1, which was measured using a chimeric myosin. We also successfully crystallized myosin XI. Structural comparison of various myosins and mutation experiments of actin-binding regions suggests that the central regions that define the fast movement of
Chara myosin XI are the actin-binding sites.
Cytoplasmic streaming with extremely high velocity (∼70 μm s
−1) occurs in cells of the characean algae (
Chara). Because cytoplasmic streaming is caused by myosin XI, it has been suggested that a myosin XI with a velocity of 70 μm s
−1, the fastest myosin measured so far, exists in
Chara cells. However, the velocity of the previously cloned
Chara corallina myosin XI (
CcXI) was about 20 μm s
−1, one-third of the cytoplasmic streaming velocity in
Chara. Recently, the genome sequence of
Chara braunii has been published, revealing that this alga has four myosin XI genes. We cloned these four myosin XI (
CbXI-1, 2, 3, and 4) and measured their velocities. While the velocities of
CbXI-3 and
CbXI-4 motor domains (MDs) were similar to that of
CcXI MD, the velocities of
CbXI-1 and
CbXI-2 MDs were 3.2 times and 2.8 times faster than that of
CcXI MD, respectively. The velocity of chimeric
CbXI-1, a functional, full-length
CbXI-1 construct, was 60 μm s
−1. These results suggest that
CbXI-1 and
CbXI-2 would be the main contributors to cytoplasmic streaming in
Chara cells and show that these myosins are ultrafast myosins with a velocity 10 times faster than fast skeletal muscle myosins in animals. We also report an atomic structure (2.8-Å resolution) of myosin XI using X-ray crystallography. Based on this crystal structure and the recently published cryo-electron microscopy structure of acto-myosin XI at low resolution (4.3-Å), it appears that the actin-binding region contributes to the fast movement of
Chara myosin XI. Mutation experiments of actin-binding surface loops support this hypothesis.
