This subproject is one of many research subprojects utilizing the resources provided by a Center grant funded by NIH/NCRR. The subproject and investigator (PI) may have received primary funding from another NIH source, and thus could be represented in other CRISP entries. The institution listed is for the Center, which is not necessarily the institution for the investigator. Molecular motor proteins function in a multitude of intracellular transport processes including organelle transport, chromosome segregation, axonal transport, and signaling pathways. Motor dependent processes are critical for growth, proliferation, and the differentiation of cells and tissues. How motor function is regulated in a developmental context, and the relationship of motor dysfunction to numerous medical problems, including neurodegenerative disease and cancer, is a current focus of research activity. Our work is focused on the microtubule motor cytoplasmic dynein, and the important and unanswered questions regarding how this single motor isoform accomplishes multiple tasks.
Our aims will address three mechanisms that potentially regulate dynein targeting and/or activity. (1) First, cytoplasmic dynein contains multiple subunits. The individual subunits or subunit domains could specify where, and to what, dynein is attached.
Aim 1 includes biochemical and genetic experiments to address how the dynein light chain and light intermediate chain influence dynein functions. (2) Second, the posttranslational modification of dynein subunits might control dynein subunit activities or binding affinities. We are defining the in vivo sites of phosphorylation on dynein subunits in collaboration with Dr. John Yates and will study the significance of the target sites. The target sites on the LIC subunit will be mutated to mimic the phosphorylated or unphosphorylated state, and the phenotypes produced by transgenes that express the mutant subunits will be analyzed. (3) In a third mechanism, specific binding partners or effector proteins might mediate the targeting of the dynein motor to specific cargoes or locations. Previous studies have provided evidence that spectrin mediates the attachment of dynactin and dynein to membranes. Our collaborator, Laura Ranum (UMN), recently discovered that Spinocerebellar ataxia type 5 (SCA5), an autosomal dominant neurodegenerative disease, is caused by mutations in 2-III spectrin (SPTBN2). In Drosophila, we have shown that mutant, but not wild type, human ss-III spectrin expressed in neurons causes neurodegeneration and a rough eye phenotype. One goal is to conduct a genome-wide screen to recover modifier loci that identify novel genes in the pathogenic process. A second priority is to determine if mutations in human spectrins, and the corresponding mutations in fly ss spectrin, disrupt axonal vesicular transport in Drosophila. These studies will help to elucidate the molecular underpinnings of SCA5 pathology and neurodegenerative disease.

Public Health Relevance

We are studying the molecular basis of intracellular transport. Our work focuses on the microtubule motor cytoplasmic dynein and the mechanisms by which this motor can accomplish diverse transport tasks. Perturbations in intracellular transport are implicated in human diseases, including neurodegenerative disease, cancer, and birth defects.

Agency
National Institute of Health (NIH)
Institute
National Center for Research Resources (NCRR)
Type
Biotechnology Resource Grants (P41)
Project #
5P41RR011823-15
Application #
8171468
Study Section
Special Emphasis Panel (ZRG1-CB-H (40))
Project Start
2010-09-01
Project End
2011-08-31
Budget Start
2010-09-01
Budget End
2011-08-31
Support Year
15
Fiscal Year
2010
Total Cost
$2,425
Indirect Cost
Name
University of Washington
Department
Biochemistry
Type
Schools of Medicine
DUNS #
605799469
City
Seattle
State
WA
Country
United States
Zip Code
98195
Hollmann, Taylor; Kim, Tae Kwon; Tirloni, Lucas et al. (2018) Identification and characterization of proteins in the Amblyomma americanum tick cement cone. Int J Parasitol 48:211-224
Stieg, David C; Willis, Stephen D; Ganesan, Vidyaramanan et al. (2018) A complex molecular switch directs stress-induced cyclin C nuclear release through SCFGrr1-mediated degradation of Med13. Mol Biol Cell 29:363-375
Seixas, Adriana; Alzugaray, María Fernanda; Tirloni, Lucas et al. (2018) Expression profile of Rhipicephalus microplus vitellogenin receptor during oogenesis. Ticks Tick Borne Dis 9:72-81
Wang, Zheng; Wu, Catherine; Aslanian, Aaron et al. (2018) Defective RNA polymerase III is negatively regulated by the SUMO-Ubiquitin-Cdc48 pathway. Elife 7:
Xavier, Marina Amaral; Tirloni, Lucas; Pinto, Antônio F M et al. (2018) A proteomic insight into vitellogenesis during tick ovary maturation. Sci Rep 8:4698
Luhtala, Natalie; Aslanian, Aaron; Yates 3rd, John R et al. (2017) Secreted Glioblastoma Nanovesicles Contain Intracellular Signaling Proteins and Active Ras Incorporated in a Farnesylation-dependent Manner. J Biol Chem 292:611-628
Thakar, Sonal; Wang, Liqing; Yu, Ting et al. (2017) Evidence for opposing roles of Celsr3 and Vangl2 in glutamatergic synapse formation. Proc Natl Acad Sci U S A 114:E610-E618
Jin, Meiyan; Fuller, Gregory G; Han, Ting et al. (2017) Glycolytic Enzymes Coalesce in G Bodies under Hypoxic Stress. Cell Rep 20:895-908
Ogami, Koichi; Richard, Patricia; Chen, Yaqiong et al. (2017) An Mtr4/ZFC3H1 complex facilitates turnover of unstable nuclear RNAs to prevent their cytoplasmic transport and global translational repression. Genes Dev 31:1257-1271
Ju Lee, Hyun; Bartsch, Deniz; Xiao, Cally et al. (2017) A post-transcriptional program coordinated by CSDE1 prevents intrinsic neural differentiation of human embryonic stem cells. Nat Commun 8:1456

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