Publication:
In Vivo Characterization of Interactions Among Dynein Complex Components at Microtubule Plus Ends

Simple item page
dc.contributor.advisorWei-Lih Lee
dc.contributor.authorPlevock, Karen M
dc.contributor.departmentUniversity of Massachusetts Amherst
dc.contributor.departmentMolecular & Cellular Biology
dc.date2023-09-22T21:02:40.000
dc.date.accessioned2024-04-26T21:09:43Z
dc.date.available2010-04-15T00:00:00Z
dc.date.issued2010-01-01
dc.date.submittedMay
dc.description.abstractDynein is a minus end directed molecular motor required for numerous cellular processes during intracellular transport and mitosis. Pac1/LIS1 and Bik1/CLIP-170 are two proteins required for targeting dynein to cytoplasmic microtubule plus ends in budding yeast. The lab previously proposed a model whereby Pac1/LIS1 binds to the motor domain of dynein heavy chain, Dyn1/HC, forming a complex that interacts with the +TIP protein Bik1/CLIP170 at plus ends. This project focused on using Bimolecular Fluorescence Complementation (BiFC) to visualize protein-protein interactions among dynein pathway components in vivo. Budding yeast, Saccharomyces cerevisiae is an ideal system to manipulate dynein as it is a non-essential protein in this system. The BiFC assay fuses two non-fluorescent halves of Venus, a YFP-derivative, to proteins of interest. If an interaction between the proteins occur, the two halves are brought to close proximity and the fluorophore is reconstituted. Cells co-expressing Dyn1-VN with Pac1-VC or Bik1-VC exhibited fluorescent foci associated with microtubule plus ends, the cell cortex and spindle pole bodies (SPBs). Additionally, cells co-expressing Pac1-VC with Bik1-VN exhibited fluorescent foci associated with microtubule plus ends. Cells coexpressing Tub1-VC and Bik1-VN or Dyn1-VN have BiFC signal indicating that both interact with the microtubule directly. Pac-1 coexpressed with Tub1 had no signal above background. These data support that these three components associate at microtubule plus ends. Dyn1 and Pac1 interact with Bik1 at microtubule plus ends. Bik1 serves as a docking platform for the two, but dynein is still able to interact with microtubules, while Pac1 is not.
dc.description.degreeMaster of Science (M.S.)
dc.identifier.doihttps://doi.org/10.7275/1276562
dc.identifier.urihttps://hdl.handle.net/20.500.14394/47337
dc.relation.urlhttps://scholarworks.umass.edu/cgi/viewcontent.cgi?article=1536&context=theses&unstamped=1
dc.source.statuspublished
dc.subjectDynein Pathway
dc.subjectMitosis
dc.subjectSaccharomyces cerevisiae
dc.subjectBiMolecular Fluorescence Complementation
dc.subjectLis1/Pac1 Bik1/Clip-170
dc.subjectLife Sciences
dc.subjectMedicine and Health Sciences
dc.titleIn Vivo Characterization of Interactions Among Dynein Complex Components at Microtubule Plus Ends
dc.typecampus
dc.typearticle
dc.typethesis
digcom.contributor.authorisAuthorOfPublication|email:kplevock@gmail.com|institution:University of Massachusetts Amherst|Plevock, Karen M
digcom.date.embargo2010-04-15T00:00:00-07:00
digcom.identifiertheses/451
digcom.identifier.contextkey1276562
digcom.identifier.submissionpaththeses/451
dspace.entity.typePublication
Files
Original bundle
Now showing 1 - 1 of 1
No Thumbnail Available
Name:
KP_Masters_Thesis_revised.pdf
Size:
1.41 MB
Format:
Adobe Portable Document Format
Name: KP_Masters_Thesis_revised.pdf
Size: 1.41 MB
Kind: Adobe PDF
Collections
Masters Theses