Abstract

Nonmuscle myosin II molecules carry out a wide variety of functions within cells. There are three nonmuscle myosin II genes. We have studied the strain dependence of the ADP release mechanism in nonmuscle myosin IIA and IIB using a solution kinetic approach. The results show that positive strains accelerate the rate of ADP release whereas negative strain markedly lowers the rate. These results suggest that these myosins, and particularly nonmuscle myosin IIB, are ideally suited to act as tension bearers in cells. We are now using optical trapping nanometry to extend these studies on both nonmuscle myosin IIA and IIB. Preliminary results with nonmuscle myosin IIB indicate that it has long attachment times as would be expected from such a slow myosin. We have expressed nonmuscle myosin IIA with a GFP-tagged regulatory light chain (RLC) with particular interest in whether the myosin would still adopt the off state when this light chain is not phosphorylated. The data show that the GFP tag does not interfere with regulation of the myosin.

  Funding Agency

Agency
National Institute of Health (NIH)
Institute
National Heart, Lung, and Blood Institute (NHLBI)
Type
Investigator-Initiated Intramural Research Projects (ZIA)
Project #
1ZIAHL001786-30
Application #
7969011
Study Section
Project Start
Project End
Budget Start
Budget End
Support Year
30
Fiscal Year
2009
Total Cost
$350,639
Indirect Cost

  Institution

Name
National Heart, Lung, and Blood Institute
Department
Type
DUNS #
City
State
Country
Zip Code

  Publications

Ecsédi, Péter; Billington, Neil; Pálfy, Gyula et al. (2018) Multiple S100 protein isoforms and C-terminal phosphorylation contribute to the paralog-selective regulation of non-muscle myosin 2 filaments. J Biol Chem :
Young, Gavin; Hundt, Nikolas; Cole, Daniel et al. (2018) Quantitative mass imaging of single biological macromolecules. Science 360:423-427
Lopata, Anna; Hughes, Ruth; Tiede, Christian et al. (2018) Affimer proteins for F-actin: novel affinity reagents that label F-actin in live and fixed cells. Sci Rep 8:6572
Melli, Luca; Billington, Neil; Sun, Sara A et al. (2018) Bipolar filaments of human nonmuscle myosin 2-A and 2-B have distinct motile and mechanical properties. Elife 7:
Liu, Xiong; Billington, Neil; Shu, Shi et al. (2017) Effect of ATP and regulatory light-chain phosphorylation on the polymerization of mammalian nonmuscle myosin II. Proc Natl Acad Sci U S A 114:E6516-E6525
Chinthalapudi, Krishna; Heissler, Sarah M; Preller, Matthias et al. (2017) Mechanistic insights into the active site and allosteric communication pathways in human nonmuscle myosin-2C. Elife 6:
Bruun, Kyle; Beach, Jordan R; Heissler, Sarah M et al. (2017) Re-evaluating the roles of myosin 18A? and F-actin in determining Golgi morphology. Cytoskeleton (Hoboken) 74:205-218
Baird, Michelle A; Billington, Neil; Wang, Aibing et al. (2017) Local pulsatile contractions are an intrinsic property of the myosin 2A motor in the cortical cytoskeleton of adherent cells. Mol Biol Cell 28:240-251
Heissler, Sarah M; Sellers, James R (2016) Various Themes of Myosin Regulation. J Mol Biol 428:1927-46
Chen, Pei-Wen; Jian, Xiaoying; Heissler, Sarah M et al. (2016) The Arf GTPase-activating Protein, ASAP1, Binds Nonmuscle Myosin 2A to Control Remodeling of the Actomyosin Network. J Biol Chem 291:7517-26

Showing the most recent 10 out of 32 publications