Life Sciences & Biotechnology
Title : | Elucidating the mechanism of motor clustering and de-clustering in Saccharomyces cerevisiae. |
Area of research : | Life Sciences & Biotechnology |
Principal Investigator : | Dr. Akhilendra Pratap Bharati, Chatrapati Sahuji Maharaj Kanpur University, Uttar Pradesh |
Timeline Start Year : | 2023 |
Timeline End Year : | 2026 |
Contact info : | akhilendrapratap1986@gmail.com |
Equipments : | Ultrasonic Probe Sonicator
Laminar Air Flow (Horizontal)
Horizontal Electrophoresis
Vertical Electrophoresis
Refrigerated Centrifuge |
Details
Executive Summary : | Cin8 and Kip1 are members of the kinesin-5 subfamily, involved in microtubule (MT) cross-linking, antiparallel microtubule sliding, and microtubule destabilization for spindle assembly and spindle elongation in budding yeast. Each Kinesin-5 motor consists of an N-terminal motor domain, α-helical neck and bipolar assembly regions (BASS), and a C-terminal tail domain. The majority of kinesin-5 motors are plus-end directed, but some are minus-end directed in vitro. Recent reports suggest that Cin8 can switch directionality due to motor-motor coupling, varying ionic conditions between processive minus and plus directed movements when traveling on single MTs. In physiological conditions, it switched from minus to plus end sliding of the linked antiparallel MT but mostly maintained minus end motion between parallel MTs. The tail domain of Cin8 controls the switch in directionality, but the mechanism is still unknown. Cin8 truncation can affect clustering and de-clustering of MTs through various assays, including Cin8 motility assay, dynamic MT assay, and antiparallel MT assay. The domain responsible for the interaction with the MT can be determined using yeast two hybrid assay. Bioinformatic analysis and site-directed mutagenesis study can further investigate residues involved in interactions. Cin8 is localized to the kinetochore before Anaphase, and its role in kinetochore recruitment can be determined by deleting Cin8 and checking the recruitment of kinetochore proteins. Ipl1, an aurora kinase, plays a major role in cell division. Ncd80 complex and Dam1 play major roles in microtubule disassembly at the kinetochore. |
Total Budget (INR): | 49,17,070 |
Organizations involved