DOI: 10.1073/pnas.94.1.48. Spare the rod, spoil the regulation: Necessity for a myosin rod

Spare the rod, spoil the regulation: Necessity for a myosin rod

10.1073/pnas.94.1.48

Crossref journal-article

Proceedings of the National Academy of Sciences

Proceedings of the National Academy of Sciences (341)

Abstract

Regulation of a variety of cellular contractile events requires that vertebrate smooth and non-muscle myosin II can achieve an “off” state. To examine the role of the myosin rod in this process, we determined the minimal size at which a myosin molecule is capable of regulation via light chain phosphorylation. Expressed smooth muscle myosin subfragments with as many as 100 amino acids of the coiled-coil rod sequence did not dimerize and were active independently of phosphorylation. To test whether dimerization per se restores regulation of ATPase activity, mutants were expressed with varying lengths of rod sequence, followed by C-terminal leucine zippers to stabilize the coiled-coil. Dimerization restored partial regulation, but the presence of a length of rod approximately equal to the myosin head was necessary to achieve a completely off state. Partially regulated short dimers could be converted into fully regulated molecules by addition of native rod sequence after the zipper. These results suggest that the myosin rod mediates specific interactions with the head that are required to obtain the completely inactive state of vertebrate smooth and non-muscle myosins. If these interactions are prohibited under cellular conditions, unphosphorylated crossbridges can slowly cycle.

Bibliography

Trybus, K. M., Freyzon, Y., Faust, L. Z., & Sweeney, H. L. (1997). Spare the rod, spoil the regulation: Necessity for aâmyosinârod. Proceedings of the National Academy of Sciences, 94(1), 48â52.

Authors 4

Kathleen M. Trybus (first)
Yelena Freyzon (additional)
Lynn Z. Faust (additional)
H. Lee Sweeney (additional)

References 20 Referenced 137

10.1007/978-1-349-09814-9_8
10.1006/bbrc.1993.2422
10.1007/BF00121066
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D R O’Reilly, L K Miller, V A Luckow Baculovirus Expression Vectors: A Laboratory Manual (Freeman, New York, 1992). / Baculovirus Expression Vectors: A Laboratory Manual by O’Reilly D R (1992)
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Dates

Type	When
Created	23 years ago (July 26, 2002, 10:31 a.m.)
Deposited	3 years, 4 months ago (April 13, 2022, 2:49 p.m.)
Indexed	1 month, 2 weeks ago (July 8, 2025, 7:29 a.m.)
Issued	28 years, 7 months ago (Jan. 7, 1997)
Published	28 years, 7 months ago (Jan. 7, 1997)
Published Online	28 years, 7 months ago (Jan. 7, 1997)
Published Print	28 years, 7 months ago (Jan. 7, 1997)

Funders 0

None

BibTeX

@article{Trybus_1997, title={Spare the rod, spoil the regulation: Necessity for a myosin rod}, volume={94}, ISSN={1091-6490}, url={http://dx.doi.org/10.1073/pnas.94.1.48}, DOI={10.1073/pnas.94.1.48}, number={1}, journal={Proceedings of the National Academy of Sciences}, publisher={Proceedings of the National Academy of Sciences}, author={Trybus, Kathleen M. and Freyzon, Yelena and Faust, Lynn Z. and Sweeney, H. Lee}, year={1997}, month=jan, pages={48–52} }

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  "abstract": "<jats:p>Regulation of a variety of cellular contractile events requires\n that vertebrate smooth and non-muscle myosin II can achieve an\n \u201coff\u201d state. To examine the role of the myosin rod in this\n process, we determined the minimal size at which a myosin molecule is\n capable of regulation via light chain phosphorylation. Expressed smooth\n muscle myosin subfragments with as many as 100 amino acids of the\n coiled-coil rod sequence did not dimerize and were active independently\n of phosphorylation. To test whether dimerization per se restores\n regulation of ATPase activity, mutants were expressed with varying\n lengths of rod sequence, followed by C-terminal leucine zippers to\n stabilize the coiled-coil. Dimerization restored partial regulation,\n but the presence of a length of rod approximately equal to the myosin\n head was necessary to achieve a completely off state. Partially\n regulated short dimers could be converted into fully regulated\n molecules by addition of native rod sequence after the zipper. These\n results suggest that the myosin rod mediates specific interactions with\n the head that are required to obtain the completely inactive state of\n vertebrate smooth and non-muscle myosins. If these interactions are\n prohibited under cellular conditions, unphosphorylated crossbridges can\n slowly cycle.</jats:p>",
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