Examining kinesin processivity within a general gating framework
Kinesin-1 is a dimeric motor that transports cargo along microtubules, taking 8.2-nm steps in a hand-over-hand fashion. The ATP hydrolysis cycles of its two heads are maintained out of phase by a series of gating mechanisms, which lead to processive runs averaging ∼1 μm. A key structural element for...
| Main Authors: | , , , , , |
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| Format: | Article |
| Language: | English |
| Published: |
eLife Sciences Publications Ltd
2015-04-01
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| Series: | eLife |
| Subjects: | |
| Online Access: | https://elifesciences.org/articles/07403 |
| _version_ | 1811236885024997376 |
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| author | Johan OL Andreasson Bojan Milic Geng-Yuan Chen Nicholas R Guydosh William O Hancock Steven M Block |
| author_facet | Johan OL Andreasson Bojan Milic Geng-Yuan Chen Nicholas R Guydosh William O Hancock Steven M Block |
| author_sort | Johan OL Andreasson |
| collection | DOAJ |
| description | Kinesin-1 is a dimeric motor that transports cargo along microtubules, taking 8.2-nm steps in a hand-over-hand fashion. The ATP hydrolysis cycles of its two heads are maintained out of phase by a series of gating mechanisms, which lead to processive runs averaging ∼1 μm. A key structural element for inter-head coordination is the neck linker (NL), which connects the heads to the stalk. To examine the role of the NL in regulating stepping, we investigated NL mutants of various lengths using single-molecule optical trapping and bulk fluorescence approaches in the context of a general framework for gating. Our results show that, although inter-head tension enhances motor velocity, it is crucial neither for inter-head coordination nor for rapid rear-head release. Furthermore, cysteine-light mutants do not produce wild-type motility under load. We conclude that kinesin-1 is primarily front-head gated, and that NL length is tuned to enhance unidirectional processivity and velocity. |
| first_indexed | 2024-04-12T12:16:52Z |
| format | Article |
| id | doaj.art-83ab9678951646c7be4c7c9bd14a6bcc |
| institution | Directory Open Access Journal |
| issn | 2050-084X |
| language | English |
| last_indexed | 2024-04-12T12:16:52Z |
| publishDate | 2015-04-01 |
| publisher | eLife Sciences Publications Ltd |
| record_format | Article |
| series | eLife |
| spelling | doaj.art-83ab9678951646c7be4c7c9bd14a6bcc2022-12-22T03:33:25ZengeLife Sciences Publications LtdeLife2050-084X2015-04-01410.7554/eLife.07403Examining kinesin processivity within a general gating frameworkJohan OL Andreasson0Bojan Milic1Geng-Yuan Chen2Nicholas R Guydosh3William O Hancock4Steven M Block5Department of Physics, Stanford University, Stanford, United StatesDepartment of Biology, Stanford University, Stanford, United StatesDepartment of Biomedical Engineering, Pennsylvania State University, University Park, United StatesBiophysics Program, Stanford University, Stanford, United StatesDepartment of Biomedical Engineering, Pennsylvania State University, University Park, United StatesDepartment of Biology, Stanford University, Stanford, United States; Department of Applied Physics, Stanford University, Stanford, United StatesKinesin-1 is a dimeric motor that transports cargo along microtubules, taking 8.2-nm steps in a hand-over-hand fashion. The ATP hydrolysis cycles of its two heads are maintained out of phase by a series of gating mechanisms, which lead to processive runs averaging ∼1 μm. A key structural element for inter-head coordination is the neck linker (NL), which connects the heads to the stalk. To examine the role of the NL in regulating stepping, we investigated NL mutants of various lengths using single-molecule optical trapping and bulk fluorescence approaches in the context of a general framework for gating. Our results show that, although inter-head tension enhances motor velocity, it is crucial neither for inter-head coordination nor for rapid rear-head release. Furthermore, cysteine-light mutants do not produce wild-type motility under load. We conclude that kinesin-1 is primarily front-head gated, and that NL length is tuned to enhance unidirectional processivity and velocity.https://elifesciences.org/articles/07403optical trapmolecular motorsingle molecule |
| spellingShingle | Johan OL Andreasson Bojan Milic Geng-Yuan Chen Nicholas R Guydosh William O Hancock Steven M Block Examining kinesin processivity within a general gating framework eLife optical trap molecular motor single molecule |
| title | Examining kinesin processivity within a general gating framework |
| title_full | Examining kinesin processivity within a general gating framework |
| title_fullStr | Examining kinesin processivity within a general gating framework |
| title_full_unstemmed | Examining kinesin processivity within a general gating framework |
| title_short | Examining kinesin processivity within a general gating framework |
| title_sort | examining kinesin processivity within a general gating framework |
| topic | optical trap molecular motor single molecule |
| url | https://elifesciences.org/articles/07403 |
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