During development, the actomyosin cytoskeleton serves as a machine to generate contractile force. Sculpting tissues requires this force-generating machine to be precisely regulated in both space and time. The small GTPase RhoA coordinates both myosin motor activation and actin filament assembly, to promote contractility. RhoA is pivotal in regulating actomyosin contractility in numerous developmental contexts, including epithelium folding. One common feature of actomyosin contractility, during both Drosophila gastrulation and Xenopus neural tube closure, is that actomyosin exhibits oscillatory dynamics. Actomyosin dynamics are critical for epithelial folding in both Drosophila gastrulation and the Xenopus neural tube. In Drosophila, these dynamics require negative regulation of RhoA via a GTPase Activating Protein (GAP).
Aim 1 of this proposal aims to understand how this Drosophila GAP is regulated to promote actomyosin dynamics and tissue folding. In addition, we have discovered a novel oscillatory behavior for an upstream guanine nucleotide exchange factor (GEF), which activates RhoA.
Aim 2 of this proposal seeks to understand the mechanism that regulates this Drosophila GEF and, thus, paces actomyosin contractility. Overall, these two aims will determine how the partnership of a specific GEF and GAP precisely regulate RhoA signaling to promote tissue folding. Because Rho-family GTPases are critical regulators of tissue structure and there are many GEFs and GAPs for Rho-family GTPases, Aim 3 will determine the regulatory logic that influences GTPase activity for other morphogenetic processes in Drosophila. First, we will examine a change in tissue integrity, epithelial-to-mesenchymal transition (EMT). We will determine mechanisms by which GTPase regulation promotes this transition and, thus, loss of epithelial integrity. Finally, we will perform a screen to identify additional combinations of GEFs and GAPs that regulate other tissue shape changes. Our overall goal is to better define how Rho-family GEFs, GAPs, and their cognate GTPases function to precisely control tissue shape and integrity.

Public Health Relevance

Tissue shape is critical to embryonic development, and defects in tissue folding lead to birth defects, such as spina bifida. We aim to identify the underlying rules that govern tissue folding, using the model system Drosophila melanogaster (fruit fly). These studies will provide insight into the regulation of a pivotal signaling molecule, which has relevance to many processes involving changes in tissue shape or integrity.

Agency
National Institute of Health (NIH)
Institute
National Institute of General Medical Sciences (NIGMS)
Type
Research Project (R01)
Project #
1R01GM125646-01
Application #
9420921
Study Section
Development - 2 Study Section (DEV2)
Program Officer
Gibbs, Kenneth D
Project Start
2018-01-01
Project End
2021-12-31
Budget Start
2018-01-01
Budget End
2018-12-31
Support Year
1
Fiscal Year
2018
Total Cost
Indirect Cost
Name
Massachusetts Institute of Technology
Department
Biology
Type
Schools of Arts and Sciences
DUNS #
001425594
City
Cambridge
State
MA
Country
United States
Zip Code
02142