Successful Problem Solving in Genetics Varies Based on Question Content.

Problem solving is a critical skill in many disciplines but is often a challenge for students to learn. To examine the processes both students and experts undertake to solve constructed-response problems in genetics, we collected the written step-by-step procedures individuals used to solve problems in four different content areas. We developed a set of codes to describe each cognitive and metacognitive process and then used these codes to describe more than 1800 student and 149 expert answers. We found that students used some processes differently depending on the content of the question, but reasoning was consistently predictive of successful problem solving across all content areas. We also confirmed previous findings that the metacognitive processes of planning and checking were more common in expert answers than student answers. We provide suggestions for instructors on how to highlight key procedures based on each specific genetics content area that can help students learn the skill of problem solving.

Characterizing Students' Ideas about the Effects of a Mutation in a Noncoding Region of DNA.

Understanding student ideas in large-enrollment biology courses can be challenging, because easy-to-administer multiple-choice questions frequently do not fully capture the diversity of student ideas. As part of the Automated Analysis of Constructed Responses (AACR) project, we designed a question prompting students to describe the possible effects of a mutation in a noncoding region of DNA. We characterized answers from 1127 students enrolled in eight different large-enrollment introductory biology courses at three different institutions over five semesters and generated an analytic scoring system containing three categories of correct ideas and five categories of incorrect ideas. We iteratively developed a computer model for scoring student answers and tested the model before and after implementing an instructional activity designed to help a new set of students explore this concept. After completing a targeted activity and re-answering the question, students showed improvement from preassessment, with 64% of students in incorrect and 67% of students in partially incorrect (mixed) categories shifting to correct ideas only. This question, computer-scoring model, and instructional activity can now be reliably used by other instructors to better understand and characterize student ideas on the effects of mutations outside a gene-coding region.

Opposing Kinesin and Myosin-I Motors Drive Membrane Deformation and Tubulation along Engineered Cytoskeletal Networks.

Microtubule and actin filament molecular motors such as kinesin-1 and myosin-Ic (Myo1c) transport and remodel membrane-bound vesicles; however, it is unclear how they coordinate to accomplish these tasks. We introduced kinesin-1- and Myo1c-bound giant unilamellar vesicles (GUVs) into a micropatterned in vitro cytoskeletal matrix modeled after the subcellular architecture where vesicular sorting and membrane remodeling are observed. This array was composed of sparse microtubules intersecting regions dense with actin filaments, and revealed that Myo1c-dependent tethering of GUVs enabled kinesin-1-driven membrane deformation and tubulation. Membrane remodeling at actin/microtubule intersections was modulated by lipid composition and the addition of the Bin-Amphiphysin-Rvs-domain (BAR-domain) proteins endophilin or FCH-domain-only (FCHo). Myo1c not only tethered microtubule-transported cargo, but also transported, deformed, and tubulated GUVs along actin filaments in a lipid-composition- and BAR-protein-responsive manner. These results suggest a mechanism for actin-based involvement in vesicular transport and remodeling of intracellular membranes, and implicate lipid composition as a key factor in determining whether vesicles will undergo transport, deformation, or tubulation driven by opposing actin and microtubule motors and BAR-domain proteins.

Myosin-I molecular motors at a glance.

Myosin-I molecular motors are proposed to play various cellular roles related to membrane dynamics and trafficking. In this Cell Science at a Glance article and the accompanying poster, we review and illustrate the proposed cellular functions of metazoan myosin-I molecular motors by examining the structural, biochemical, mechanical and cell biological evidence for their proposed molecular roles. We highlight evidence for the roles of myosin-I isoforms in regulating membrane tension and actin architecture, powering plasma membrane and organelle deformation, participating in membrane trafficking, and functioning as a tension-sensitive dock or tether. Collectively, myosin-I motors have been implicated in increasingly complex cellular phenomena, yet how a single isoform accomplishes multiple types of molecular functions is still an active area of investigation. To fully understand the underlying physiology, it is now essential to piece together different approaches of biological investigation. This article will appeal to investigators who study immunology, metabolic diseases, endosomal trafficking, cell motility, cancer and kidney disease, and to those who are interested in how cellular membranes are coupled to the underlying actin cytoskeleton in a variety of different applications.

Control of the initiation and termination of kinesin-1-driven transport by myosin-Ic and nonmuscle tropomyosin.

Intracellular transport is largely driven by processive microtubule- and actin-based molecular motors. Nonprocessive motors have also been localized to trafficking cargos, but their roles are not well understood. Myosin-Ic (Myo1c), a nonprocessive actin motor, functions in a variety of exocytic events, although the underlying mechanisms are not yet clear. To investigate the interplay between myosin-I and the canonical long-distance transport motor kinesin-1, we attached both motor types to lipid membrane-coated bead cargo, using an attachment strategy that allows motors to actively reorganize within the membrane in response to the local cytoskeletal environment. We compared the motility of kinesin-1-driven cargos in the absence and presence of Myo1c at engineered actin/microtubule intersections. We found that Myo1c significantly increases the frequency of kinesin-1-driven microtubule-based runs that begin at actin/microtubule intersections. Myo1c also regulates the termination of processive runs. Beads with both motors bound have a significantly higher probability of pausing at actin/microtubule intersections, remaining tethered for an average of 20 s, with some pauses lasting longer than 200 s. The actin-binding protein nonmuscle tropomyosin (Tm) provides spatially specific regulation of interactions between myosin motors and actin filaments in vivo; in the crossed-filament in vitro assay, we found that Tm2-actin abolishes Myo1c-specific effects on both run initiation and run termination. Together, these observations suggest Myo1c is important for the selective initiation and termination of kinesin-1-driven runs along microtubules at specific actin filament populations within the cell.