scholarly journals Concerted action of kinesins KIF5B and KIF13B promotes efficient secretory vesicle transport to microtubule plus ends

eLife ◽  
2020 ◽  
Vol 9 ◽  
Author(s):  
Andrea Serra-Marques ◽  
Maud Martin ◽  
Eugene A Katrukha ◽  
Ilya Grigoriev ◽  
Cathelijn AE Peeters ◽  
...  
Keyword(s):  
Golgi Apparatus ◽  
Intracellular Transport ◽  
Dominant Role ◽  
Secretory Vesicle ◽  
Vesicle Transport ◽  
Reverse Direction ◽  
Cell Periphery ◽  
Secretory Vesicles ◽  
Concerted Action ◽  
Directed Transport

Intracellular transport relies on multiple kinesins, but it is poorly understood which kinesins are present on particular cargos, what their contributions are and whether they act simultaneously on the same cargo. Here, we show that Rab6-positive secretory vesicles are transported from the Golgi apparatus to the cell periphery by kinesin-1 KIF5B and kinesin-3 KIF13B, which determine the location of secretion events. KIF5B plays a dominant role, whereas KIF13B helps Rab6 vesicles to reach freshly polymerized microtubule ends, to which KIF5B binds poorly, likely because its cofactors, MAP7-family proteins, are slow in populating these ends. Sub-pixel localization demonstrated that during microtubule plus-end directed transport, both kinesins localize to the vesicle front and can be engaged on the same vesicle. When vesicles reverse direction, KIF13B relocates to the middle of the vesicle, while KIF5B shifts to the back, suggesting that KIF5B but not KIF13B undergoes a tug-of-war with a minus-end directed motor.

scholarly journals Concerted action of kinesins KIF5B and KIF13B promotes efficient secretory vesicle transport to microtubule plus ends

2020 ◽  
Author(s):  
Andrea Serra-Marques ◽  
Maud Martin ◽  
Eugene A. Katrukha ◽  
Ilya Grigoriev ◽  
Cathelijn A.E. Peeters ◽  
...  
Keyword(s):  
Golgi Apparatus ◽  
Intracellular Transport ◽  
Dominant Role ◽  
Secretory Vesicle ◽  
Vesicle Transport ◽  
Reverse Direction ◽  
Cell Periphery ◽  
Secretory Vesicles ◽  
Concerted Action ◽  
Directed Transport

AbstractIntracellular transport relies on multiple kinesins, but it is poorly understood which kinesins are present on particular cargos, what their contributions are and whether they act simultaneously on the same cargo. Here, we show that Rab6-positive secretory vesicles are transported from the Golgi apparatus to the cell periphery by kinesin-1 KIF5B and kinesin-3 KIF13B, which determine the location of secretion events. KIF5B plays a dominant role, whereas KIF13B helps Rab6 vesicles to reach freshly polymerized microtubule ends, to which KIF5B binds poorly, likely because its cofactors, MAP7-family proteins, are slow in populating these ends. Sub-pixel localization demonstrated that during microtubule plus-end directed transport, both kinesins localize to the vesicle front and can be engaged on the same vesicle. When vesicles reverse direction, KIF13B relocates to the middle of the vesicle, while KIF5B shifts to the back, suggesting that KIF5B but not KIF13B undergoes a tug-of-war with a minus-end directed motor.

scholarly journals Yeast Rgd3 is a phospho-regulated F-BAR–containing RhoGAP involved in the regulation of Rho3 distribution and cell morphology

2020 ◽  
Vol 31 (23) ◽  
pp. 2570-2582
Author(s):  
Robert M. Gingras ◽  
Kyaw Myo Lwin ◽  
Abigail M. Miller ◽  
Anthony Bretscher
Keyword(s):  
Cell Polarity ◽  
Temperature Sensitivity ◽  
Cell Morphology ◽  
Budding Yeast ◽  
Secretory Vesicle ◽  
Vesicle Transport ◽  
Secretory Vesicles

RGD3 ( YHR182w) is a novel RhoGAP in budding yeast whose overexpression suppresses the temperature sensitivity of myo2 smy1 mutants defective in secretory vesicle transport. We show that Rgd3 localizes to polarized vesicles distinct from constitutive secretory vesicles and contributes to cell polarity through its GAP activity on Rho3.

Golgi Apparatus Secretion of Vacuolar Substances in Maize Root Cells

1976 ◽  
Vol 34 ◽  
pp. 44-45
Author(s):  
H. H. Mollenhauer ◽  
J. E. Hanson
Keyword(s):  
Cell Wall ◽  
Golgi Apparatus ◽  
Mammalian Cells ◽  
Secretory Vesicle ◽  
General Mechanism ◽  
Secretory Vesicles ◽  
Extracellular Medium ◽  
Root Cells ◽  
Ultrastructural Studies ◽  
Secretory Products

It has long been known that secretory vesicles from the Golgi apparatus discharge their contents through the plasmalemma by a process of membrane fusion called exocytosis (2,3). Ultrastructural studies have shown that the membrane of the secretory vesicle fuses with the plasmalemma, opens up at the site of fusion, and then discharges the vesicle product to the extracellular medium (i.e., to the lumen or cell wall) (2,3). Exocytosis is recognized as a widely occurring, and possibly general, mechanism for the discharge of macromolecular secretory products (2,3). A similar mechanism is also operable for the transfer of Golgi apparatus product into the acrosome (a lysosome) of mammalian cells (4).

scholarly journals Author response: Concerted action of kinesins KIF5B and KIF13B promotes efficient secretory vesicle transport to microtubule plus ends

2020 ◽  
Author(s):  
Andrea Serra-Marques ◽  
Maud Martin ◽  
Eugene A Katrukha ◽  
Ilya Grigoriev ◽  
Cathelijn AE Peeters ◽  
...  
Keyword(s):  
Secretory Vesicle ◽  
Vesicle Transport ◽  
Author Response ◽  
Concerted Action

scholarly journals Polarized Growth in Budding Yeast in the Absence of a Localized Formin

2009 ◽  
Vol 20 (10) ◽  
pp. 2540-2548 ◽  
Author(s):  
Lina Gao ◽  
Anthony Bretscher
Keyword(s):  
Budding Yeast ◽  
Myosin Ii ◽  
Secretory Vesicle ◽  
Secretory Vesicles ◽  
Polarized Growth ◽  
Directed Transport ◽  
Bud Neck ◽  
Actin Cables ◽  
In Cells

Polarity is achieved partly through the localized assembly of the cytoskeleton. During growth in budding yeast, the bud cortex and neck localized formins Bni1p and Bnr1p nucleate and assemble actin cables that extend along the bud-mother axis, providing tracks for secretory vesicle delivery. Localized formins are believed to determine the location and polarity of cables, hence growth. However, yeast expressing the nonlocalized actin nucleating/assembly formin homology (FH) 1-FH2 domains of Bnr1p or Bni1p as the sole formin grow well. Although cables are significantly disorganized, analysis of directed transport of secretory vesicles is still biased toward the bud, reflecting a bias in correctly oriented cables, thereby permitting polarized growth. Myosin II, localized at the bud neck, contributes to polarized growth as a mutant unable to interact with F-actin further compromises growth in cells with an unlocalized formin but not with a localized formin. Our results show that multiple mechanisms contribute to cable orientation and polarized growth, with localized formins and myosin II being two major contributors.

Synaptotagmin-1: A Multi-Functional Protein that Mediates Vesicle Docking, Priming, and Fusion

2021 ◽  
Vol 22 ◽  
Author(s):  
Najeeb Ullah ◽  
Ezzouhra El Maaiden ◽  
Md. Sahab Uddin ◽  
Ghulam Md Ashraf
Keyword(s):  
Plasma Membrane ◽  
Secretory Granules ◽  
Secretory Vesicle ◽  
Neuroendocrine Cells ◽  
Snare Complex ◽  
Secretory Vesicles ◽  
Functional Protein ◽  
Vesicle Docking ◽  
Synaptotagmin 1

: The fusion of secretory vesicles with the plasma membrane depends on the assembly of v-SNAREs (VAMP2/synaptobrevin2) and t-SNAREs (SNAP25/syntaxin1) into the SNARE complex. Vesicles go through several upstream steps, referred to as docking and priming, to gain fusion competence. The vesicular protein synaptotagmin-1 (Syt-1) is the principal Ca2+ sensor for fusion in several central nervous system neurons and neuroendocrine cells and part of the docking complex for secretory granules. Syt-1 binds to the acceptor complex such as synaxin1, SNAP-25 on the plasma membrane to facilitate secretory vesicle docking, and upon Ca2+-influx promotes vesicle fusion. This review assesses the role of the Syt-1 protein involved in the secretory vesicle docking, priming, and fusion.

scholarly journals The Cooh-Terminal Domain of Myo2p, a Yeast Myosin V, Has a Direct Role in Secretory Vesicle Targeting

1999 ◽  
Vol 147 (4) ◽  
pp. 791-808 ◽  
Author(s):  
Daniel Schott ◽  
Jackson Ho ◽  
David Pruyne ◽  
Anthony Bretscher
Keyword(s):  
Secretory Vesicle ◽  
Restrictive Temperature ◽  
Secretory Vesicles ◽  
Tail Domain ◽  
Myosin V ◽  
Direct Role ◽  
Rab Protein ◽  
Polarized Delivery ◽  
Actin Cables ◽  
Type V

MYO2 encodes a type V myosin heavy chain needed for the targeting of vacuoles and secretory vesicles to the growing bud of yeast. Here we describe new myo2 alleles containing conditional lethal mutations in the COOH-terminal tail domain. Within 5 min of shifting to the restrictive temperature, the polarized distribution of secretory vesicles is abolished without affecting the distribution of actin or the mutant Myo2p, showing that the tail has a direct role in vesicle targeting. We also show that the actin cable–dependent translocation of Myo2p to growth sites does not require secretory vesicle cargo. Although a fusion protein containing the Myo2p tail also concentrates at growth sites, this accumulation depends on the polarized delivery of secretory vesicles, implying that the Myo2p tail binds to secretory vesicles. Most of the new mutations alter a region of the Myo2p tail conserved with vertebrate myosin Vs but divergent from Myo4p, the myosin V involved in mRNA transport, and genetic data suggest that the tail interacts with Smy1p, a kinesin homologue, and Sec4p, a vesicle-associated Rab protein. The data support a model in which the Myo2p tail tethers secretory vesicles, and the motor transports them down polarized actin cables to the site of exocytosis.

scholarly journals Visualization of Intracellular Transport of Vesicular Stomatitis Virus Nucleocapsids in Living Cells

2006 ◽  
Vol 80 (13) ◽  
pp. 6368-6377 ◽  
Author(s):  
Subash C. Das ◽  
Debasis Nayak ◽  
You Zhou ◽  
Asit K. Pattnaik
Keyword(s):  
Vesicular Stomatitis Virus ◽  
Protein Function ◽  
Intracellular Transport ◽  
Fluorescent Protein ◽  
De Novo ◽  
Virus Production ◽  
Hinge Region ◽  
Cell Periphery ◽  
P Protein ◽  
Vesicular Stomatitis

ABSTRACT The phosphoprotein (P) of vesicular stomatitis virus (VSV) is a subunit of the viral RNA polymerase. In previous studies, we demonstrated that insertion of 19 amino acids in the hinge region of the protein had no significant effect on P protein function. In the present study, we inserted full-length enhanced green fluorescent protein (eGFP) in frame into the hinge region of P and show that the fusion protein (PeGFP) is functional in viral genome transcription and replication, albeit with reduced activity. A recombinant vesicular stomatitis virus encoding PeGFP in place of the P protein (VSV-PeGFP), which possessed reduced growth kinetics compared to the wild-type VSV, was recovered. Using the recombinant VSV-PeGFP, we show that the viral replication proteins and the de novo-synthesized RNA colocalize to sites throughout the cytoplasm, indicating that replication and transcription are not confined to any particular region of the cytoplasm. Real-time imaging of the cells infected with the eGFP-tagged virus revealed that, following synthesis, the nucleocapsids are transported toward the cell periphery via a microtubule (MT)-mediated process, and the nucleocapsids were seen to be closely associated with mitochondria. Treatment of cells with nocodazole or Colcemid, drugs known to inhibit MT polymerization, resulted in accumulation of the nucleocapsids around the nucleus and also led to inhibition of infectious-virus production. These findings are compatible with a model in which the progeny viral nucleocapsids are transported toward the cell periphery by MT and the transport may be facilitated by mitochondria.

Sign in / Sign up

Export Citation Format

Share Document