Kinesin-3 motors are fine-tuned at the molecular level to endow distinct mechanical outputs
Abstract Background Kinesin-3 family motors drive diverse cellular processes and have significant clinical importance. The ATPase cycle is integral to the processive motility of kinesin motors to drive long-distance intracellular transport. Our previous work has demonstrated that kinesin-3 motors ar...
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| Format: | Article |
| Language: | English |
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BMC
2022-08-01
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| Series: | BMC Biology |
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| Online Access: | https://doi.org/10.1186/s12915-022-01370-8 |
| _version_ | 1818499058611781632 |
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| author | Pushpanjali Soppina Nishaben Patel Dipeshwari J. Shewale Ashim Rai Sivaraj Sivaramakrishnan Pradeep K. Naik Virupakshi Soppina |
| author_facet | Pushpanjali Soppina Nishaben Patel Dipeshwari J. Shewale Ashim Rai Sivaraj Sivaramakrishnan Pradeep K. Naik Virupakshi Soppina |
| author_sort | Pushpanjali Soppina |
| collection | DOAJ |
| description | Abstract Background Kinesin-3 family motors drive diverse cellular processes and have significant clinical importance. The ATPase cycle is integral to the processive motility of kinesin motors to drive long-distance intracellular transport. Our previous work has demonstrated that kinesin-3 motors are fast and superprocessive with high microtubule affinity. However, chemomechanics of these motors remain poorly understood. Results We purified kinesin-3 motors using the Sf9-baculovirus expression system and demonstrated that their motility properties are on par with the motors expressed in mammalian cells. Using biochemical analysis, we show for the first time that kinesin-3 motors exhibited high ATP turnover rates, which is 1.3- to threefold higher compared to the well-studied kinesin-1 motor. Remarkably, these ATPase rates correlate to their stepping rate, suggesting a tight coupling between chemical and mechanical cycles. Intriguingly, kinesin-3 velocities (KIF1A > KIF13A > KIF13B > KIF16B) show an inverse correlation with their microtubule-binding affinities (KIF1A < KIF13A < KIF13B < KIF16B). We demonstrate that this differential microtubule-binding affinity is largely contributed by the positively charged residues in loop8 of the kinesin-3 motor domain. Furthermore, microtubule gliding and cellular expression studies displayed significant microtubule bending that is influenced by the positively charged insert in the motor domain, K-loop, a hallmark of kinesin-3 family. Conclusions Together, we propose that a fine balance between the rate of ATP hydrolysis and microtubule affinity endows kinesin-3 motors with distinct mechanical outputs. The K-loop, a positively charged insert in the loop12 of the kinesin-3 motor domain promotes microtubule bending, an interesting phenomenon often observed in cells, which requires further investigation to understand its cellular and physiological significance. |
| first_indexed | 2024-12-10T20:24:06Z |
| format | Article |
| id | doaj.art-ad9f4edd701d45a5b0c8b228b8328983 |
| institution | Directory Open Access Journal |
| issn | 1741-7007 |
| language | English |
| last_indexed | 2024-12-10T20:24:06Z |
| publishDate | 2022-08-01 |
| publisher | BMC |
| record_format | Article |
| series | BMC Biology |
| spelling | doaj.art-ad9f4edd701d45a5b0c8b228b83289832022-12-22T01:34:56ZengBMCBMC Biology1741-70072022-08-0120112310.1186/s12915-022-01370-8Kinesin-3 motors are fine-tuned at the molecular level to endow distinct mechanical outputsPushpanjali Soppina0Nishaben Patel1Dipeshwari J. Shewale2Ashim Rai3Sivaraj Sivaramakrishnan4Pradeep K. Naik5Virupakshi Soppina6Discipline of Biological Engineering, Indian Institute of Technology GandhinagarDiscipline of Biological Engineering, Indian Institute of Technology GandhinagarDiscipline of Biological Engineering, Indian Institute of Technology GandhinagarDepartment of Genetics, Cell Biology and Development, University of MinnesotaDepartment of Genetics, Cell Biology and Development, University of MinnesotaDepartment of Biotechnology and Bioinformatics, Sambalpur UniversityDiscipline of Biological Engineering, Indian Institute of Technology GandhinagarAbstract Background Kinesin-3 family motors drive diverse cellular processes and have significant clinical importance. The ATPase cycle is integral to the processive motility of kinesin motors to drive long-distance intracellular transport. Our previous work has demonstrated that kinesin-3 motors are fast and superprocessive with high microtubule affinity. However, chemomechanics of these motors remain poorly understood. Results We purified kinesin-3 motors using the Sf9-baculovirus expression system and demonstrated that their motility properties are on par with the motors expressed in mammalian cells. Using biochemical analysis, we show for the first time that kinesin-3 motors exhibited high ATP turnover rates, which is 1.3- to threefold higher compared to the well-studied kinesin-1 motor. Remarkably, these ATPase rates correlate to their stepping rate, suggesting a tight coupling between chemical and mechanical cycles. Intriguingly, kinesin-3 velocities (KIF1A > KIF13A > KIF13B > KIF16B) show an inverse correlation with their microtubule-binding affinities (KIF1A < KIF13A < KIF13B < KIF16B). We demonstrate that this differential microtubule-binding affinity is largely contributed by the positively charged residues in loop8 of the kinesin-3 motor domain. Furthermore, microtubule gliding and cellular expression studies displayed significant microtubule bending that is influenced by the positively charged insert in the motor domain, K-loop, a hallmark of kinesin-3 family. Conclusions Together, we propose that a fine balance between the rate of ATP hydrolysis and microtubule affinity endows kinesin-3 motors with distinct mechanical outputs. The K-loop, a positively charged insert in the loop12 of the kinesin-3 motor domain promotes microtubule bending, an interesting phenomenon often observed in cells, which requires further investigation to understand its cellular and physiological significance.https://doi.org/10.1186/s12915-022-01370-8ChemomechanicalKinesin-3Microtubule bendingBaculovirusSuperprocessiveATPases |
| spellingShingle | Pushpanjali Soppina Nishaben Patel Dipeshwari J. Shewale Ashim Rai Sivaraj Sivaramakrishnan Pradeep K. Naik Virupakshi Soppina Kinesin-3 motors are fine-tuned at the molecular level to endow distinct mechanical outputs BMC Biology Chemomechanical Kinesin-3 Microtubule bending Baculovirus Superprocessive ATPases |
| title | Kinesin-3 motors are fine-tuned at the molecular level to endow distinct mechanical outputs |
| title_full | Kinesin-3 motors are fine-tuned at the molecular level to endow distinct mechanical outputs |
| title_fullStr | Kinesin-3 motors are fine-tuned at the molecular level to endow distinct mechanical outputs |
| title_full_unstemmed | Kinesin-3 motors are fine-tuned at the molecular level to endow distinct mechanical outputs |
| title_short | Kinesin-3 motors are fine-tuned at the molecular level to endow distinct mechanical outputs |
| title_sort | kinesin 3 motors are fine tuned at the molecular level to endow distinct mechanical outputs |
| topic | Chemomechanical Kinesin-3 Microtubule bending Baculovirus Superprocessive ATPases |
| url | https://doi.org/10.1186/s12915-022-01370-8 |
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