551-1417-00L In Vivo Cryo-EM Analysis of Dynein Motor Proteins
|Autumn Semester 2020
|yearly recurring course
|Language of instruction
|Number of participants limited to 5.
The enrolment is done by the D-BIOL study administration.
General safety regulations for all block courses:
-Whenever possible the distance rules have to be respected
-All students have to wear masks throughout the course (keep reserve masks ready)
-The installation and activation of the Swiss Covid-App is highly encouraged
-Any additional rules for individual courses have to be respected
-Students showing any COVID-19 symptoms are not allowed to enter ETH buildings and have to inform the course responsible
|In Vivo Cryo-EM Analysis of Dynein Motor Proteins
Permission from lecturers required for all students.
Block course in the 4th quarter of the autumn semester
Place: Paul Scherrer Institut, Villigen
|Motor proteins convert chemical energy into mechanical motion. In this block course, we study dynein motor proteins in cilia. Dynein causes conformational change upon ATP hydrolysis and finally generate ciliary bending motion. Participants will analyze cryo-EM data of cilia and visualize in vivo 3D structure of dynein to learn how motor proteins function in the cell.
|The goal of this course is to be familiar with structural biology techniques of cryo-electron tomography and single particle cryo-EM studies on motor proteins. The main focus is 3D image analysis of cryo-EM datasets acquired by highest-end microscopes. Participants will learn structure-function relationship at various scales: how the conformational change of motor proteins causes mechanical force and generates cellular motility.
|Motor proteins, such as dynein, myosin and kinesin, hydrolyze ATP to ADP and phosphate to convert chemical energy to mechanical motion. Their function is essential for intracellular transport, muscle contraction and other cellular motility as well as cell division. Motor proteins have been major targets of biophysical studies. There exist questions from atomic to tissue levels – how ATP hydrolysis causes conformational change of motor proteins; how their motion is regulated by calcium, phosphorylation and other factors; how motions of multiple motor proteins are coordinated to generate cellular motility. Structural biology has been playing central roles to answer these questions. X-ray crystallography and single particle cryo-EM address structural analysis at atomic resolution and try to reveal molecular mechanism of conformational change. Cryo-electron tomography analyze localization and 3D structure of motor proteins in the cell to explain how motions of molecular motors happen in the context of cellular environment and are integrated into cellular motion.
In this course, we study dyneins in cilia. Cilia are force-generating organelles, made by nine microtubules and thousands of dyneins. Dynein hydrolyzes ATP and undergoes conformational change, generating linear motion with respect to the microtubule. As a whole system, cilia integrate motions of these dyneins and orchestrate beating motion. To explain ciliary motion at molecular level, we need to know dynein conformational change in the cellular context. Cryo-electron tomography is recently developed technique to study molecular structures in vivo and therefore a suitable method to study dynein in cilia. Recently spatial resolution of these cryo-EM techniques was dramatically improved, driven by development of new types of detectors and electron optics.
The participants of this course will learn a program to analyze cryo-electron tomography and single particle cryo-EM data, acquired by highest-end electron microscopes and detectors in ETH and other places, and reconstruct 3D structure (tomogram) of cilia from various organisms (from green algae to human). They will further learn a program to study molecular structures from these tomograms (called subtomogram averaging) and apply it to reconstruct high-resolution 3D structure of dyneins, microtubules and regulatory proteins. This practical course is therefore mainly computational, but we will also provide students a chance of cilia preparation from green algae, cryo-EM data collection using an electron microscope in PSI and site-visit of highest-end electron microscope facility in ETH.
|Scripts will be distributed during the course.
|An overview is given in the following review articles. Further literature will be indicated during the course.
Ishikawa (2017) “Axoneme structure from motile cilia” Cold Spring Harb. Perspect Biol. 9. doi: 10.1101/cshperspect.a028076.
Ishikawa (2017) “Cryo-electron tomography of motile cilia and flagella” Cilia 4, 3. doi: 10.1186/s13630-014-0012-7.
|Performance assessment information (valid until the course unit is held again)
|Performance assessment as a semester course
|graded semester performance
|Language of examination
|Repetition only possible after re-enrolling for the course unit.
|No public learning materials available.
|Only public learning materials are listed.
|No information on groups available.
|Permission from lecturers required for all students
|Block Courses in 4th Quarter of the Semester