Kathleen M. Trybus, Ph.D.
Dr. Kathy Trybus received her Ph.D. in 1981 from the University of Chicago in Biophysics. She next went to Brandeis University and worked in a Structural Biology lab in the Rosenstiel Basic Medical Sciences Research Center. In 1998, Dr. Trybus joined the Department faculty at University of Vermont.
A major focus of the laboratory is unconventional myosin V, a processive motor that moves cargo along actin for long distances without dissociating. Two class V myosins from budding yeast (called Myo2p and Myo4p) have captured our interest, since both had been characterized to be non-processive, despite being demonstrated cargo transporters.
Using single molecule and biochemical techniques, we recently showed that Myo2p can only move processively when it walks on actin tracks that resemble those found in the cell. In this case, yeast tropomyosin was found to be essential for the motor to move processively — an elegant demonstration of how the track can affect the motor.
Myo4p, a single-headed class V myosin in budding yeast, transports mRNA to the bud tip. We showed that an oligomeric adapter protein that links the motor to the cargo (She2p) recruits two motors to form a processive complex. Addition of mRNA cargo greatly stabilizes the complex, so that only motors with cargo can move processively. We are altering the mRNA cargo to understand why localizing mRNAs have multiple “zipcode” elements.
Vertebrate myosin Va undergoes a folded (inactive) to extended (active) conformational transition. We are investigating if cargo binding is sufficient to activate the motor, using myoVa-melanophilin-Rab27a-melanosome as a model system.
Another major focus is to understand the molecular mechanisms by which point mutations in smooth muscle actin (ACTA2) lead to vascular disease. These studies are made possible by our ability to express homogeneous wild-type and mutant vertebrate actins in the baculovirus/insect cell expression system. Techniques used to assess defects caused by the mutations include: a TIRF based assay that follows polymerization of single actin filaments in real time, measurement of persistence length to assess structural changes in the filament, and motility assays to quantify how fast smooth muscle myosin can move the actin filaments .
Tropomyosin is essential for Myo2p, a class V myosin in budding yeast, to move processively on actin.
Interactions between two molecular motors coupled to a DNA scaffold.
Lu H, Fagnant PM, Trybus KM (2019) Unusual dynamics of the divergent malaria parasite PfAct1 actin filament. Proc Natl Acad Sci U S A 116(41): 20418-20427.
Robert-Paganin J, Robblee JP, Auguin D, Blake TCA, Bookwalter CS, Krementsova EB, Moussaoui D, Previs MJ, Jousset G, Baum J, Trybus KM, Houdusse A (2019) Plasmodium myosin A drives parasite invasion by an atypical force generating mechanism. Nat Commun 10(1): 3286.
Chaudhary AR, Lu H, Krementsova EB, Bookwalter CS, Trybus KM, Hendricks AG (2019) MAP7 regulates organelle transport by recruiting kinesin-1 to microtubules. J Biol Chem 294(26): 10160-10171.
Lombardo AT, Nelson SR, Kennedy GG, Trybus KM, Walcott S, Warshaw DM (2019) Myosin Va transport of liposomes in three-dimensional actin networks is modulated by actin filament density, position, and polarity. Proc Natl Acad Sci U S A 116(17): 8326-8335.
Lowey S, Bretton V, Joel PB, Trybus KM, Gulick J, Robbins J, Kalganov A, Cornachione AS, Rassier DE (2018) Hypertrophic cardiomyopathy R403Q mutation in rabbit β-myosin reduces contractile function at the molecular and myofibrillar levels. Proc Natl Acad Sci U S A 115(44): 11238-11243.
Sladewski TE, Billington N, Ali MY, Bookwalter CS, Lu H, Krementsova EB, Schroer TA, Trybus KM (2018) Recruitment of two dyneins to an mRNA-dependent Bicaudal D transport complex. Elife 7: .
Tjioe M, Ryoo H, Ishitsuka Y, Ge P, Bookwalter C, Huynh W, McKenney RJ, Trybus KM, Selvin PR (2018) Magnetic Cytoskeleton Affinity Purification of Microtubule Motors Conjugated to Quantum Dots. Bioconjug Chem 29(7): 2278-2286.