Kif5B and Kifc1 Interact and Are Required for Motility and Fission of Early Endocytic Vesicles in Mouse Liver

Albert Einstein College of Medicine, New York, New York, United States
Molecular Biology of the Cell (Impact Factor: 4.47). 06/2007; 18(5):1839-49. DOI: 10.1091/mbc.E06-06-0524
Source: PubMed


Early endocytic vesicles loaded with Texas Red asialoorosomucoid were prepared from mouse liver. These vesicles bound to microtubules in vitro, and upon ATP addition, they moved bidirectionally, frequently undergoing fission into two daughter vesicles. There was no effect of vanadate (inhibitor of dynein) on motility, whereas 5'-adenylylimido-diphosphate (kinesin inhibitor) was highly inhibitory. Studies with specific antibodies confirmed that dynein was not associated with these vesicles and that Kif5B and the minus-end kinesin Kifc1 mediated their plus- and minus-end motility, respectively. More than 90% of vesicles associated with Kifc1 also contained Kif5B, and inhibition of Kifc1 with antibody resulted in enhancement of plus-end-directed motility. There was reduced vesicle fission when either Kifc1 or Kif5B activity was inhibited by antibody, indicating that the opposing forces resulting from activity of both motors are required for fission to occur. Immunoprecipitation of native Kif5B by FLAG antibody after expression of FLAG-Kifc1 in 293T cells indicates that these two motors can interact with each other. Whether they interact directly or through a complex of potential regulatory proteins will need to be clarified in future studies. However, the present study shows that coordinated activity of these kinesins is essential for motility and processing of early endocytic vesicles.

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Available from: Eustratios Bananis, Dec 31, 2015
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    • "RE Rip11/FIP5 [133] LE/lysosomes Unknown [65] [109] EE tubules; SNX4 Unknown [58] Kinesin-3 KIF1B␤ LE/lysosomes Unknown [115] [135] KIF13A RE Rab11 [132] KIF16B EE/RE PtdIns-3P [107] [109] Khc-73 (D. melanogaster) EE (Unknown) [106] Kinesin-13 KIF2␤ LE/lysosomes Unknown [116] Kinesin-14 KIFC1 EE Unknown [67] KIFC2 EE Unknown [66] Dynein/dynactin LIC1 LE/lysosomes RILP [63] LIC1/2 RE FIP3 [130] [131] LIC1 RE Rab4 [90] LIC (D. rerio) LE/lysosomes JIP3 (in neurons) [100] IC LE/lysosomes Snapin [102] LC8 EE tubules; SNX4 Kibra [129] "
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    ABSTRACT: The endocytic pathway is essential for processes that define how cells interact with their environment, including receptor signalling, cell adhesion and migration, pathogen entry, membrane protein turnover and nutrient uptake. The spatial organisation of endocytic trafficking requires motor proteins that tether membranes or transport them along the actin and microtubule cytoskeletons. Microtubules, actin filaments and motor proteins also provide force to deform and assist in the scission of membranes, thereby facilitating endosomal sorting and the generation of transport intermediates.
    Preview · Article · Jan 2014 · Seminars in Cell and Developmental Biology
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    • "This suggests that these minus-end kinesins are active in the absence of PDZK1, consistent with the relatively large amount of residual minus-end directed motility in these vesicles when dynein is inhibited with vanadate or antibody. The reason for their lack of activity in vesicles from wild-type mice is not known at this time, but could be related to interaction with kinesin-1, as we described in a previous study (Nath et al., 2007). Vesicles from PDZK1 knockout mice exhibit a different profile of motor proteins, with increased KifC3 and dynein but decreased KifC1 and Kif5B. "
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    ABSTRACT: Previous studies identified a family of organic anion transport proteins (OATPs), many of which have C-terminal PDZ binding consensus sequences. In particular, the C-terminal four amino acids of oatp1a1, a transporter on rat and mouse hepatocytes, comprise a consensus binding site for PDZK1. In PDZK1 knockout mice and in transfected cells where PDZK1 expression was knocked down, oatp1a1 accumulates in intracellular vesicles. The present study tests the hypothesis that oatp1a1 traffics to and from the cell surface in vesicles along microtubules and that PDZK1 guides recruitment of specific motors to these vesicles. Oatp1a1-containing vesicles were prepared from wild type and PDZK1 knockout mice. As seen by immunofluorescence, kinesin-1, a microtubule plus-end directed motor, was largely associated with vesicles from wild type mouse liver, while dynein, a minus-end directed motor, was largely associated with vesicles from PDZK1 knockout mouse liver. Quantification of motility on directionally marked microtubules following addition of 50 μM ATP showed that wild type vesicles moved equally towards the plus and minus ends while PDZK1 knockout vesicles moved predominantly towards the minus end, consistent with net movement towards the cell interior. These studies provide a novel mechanism by which PDZK1 regulates intracellular trafficking of oatp1a1 by recruiting specific motors to oatp1a1-containing vesicles. In the absence of PDZK1, oatp1a1-containing vesicles can't recruit kinesin-1 and associate with dynein as a predominant minus-end directed motor. Whether this is a result of direct interaction of the oatp1a1 cytoplasmic domain with dynein or with a dynein-containing protein complex remains to be established.
    Preview · Article · Oct 2013 · Drug metabolism and disposition: the biological fate of chemicals
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    • "So in other word, the function of KIFC1 in S. maindroni is similar to that in mammals . As we know, the KIFC1 protein has different roles in various tissues, such as it is essential in the fusion of vesicles in the lung of rat (Nath et al., 2007). kifc1 mRNA expressed in different tissue. "
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    ABSTRACT: In this study, we investigated the gene sequence and characteristic of kifc1 in Sepiella maindroni through PCR and RACE technology. Our research aimed particularly at the spatio-temporal expression pattern of kifc1 in the developmental testis through in situ hybridization. The particular role of kifc1 in the spermatogenesis of S. maindroni was our particular interest. Based on multiple protein sequence alignments of KIFC1 homologues, kifc1 gene from the testis of S. maindroni was identified, which consisted of 2432bp including a 2109 in-frame ORF corresponding to 703 continuous amino acids. The encoded polypeptide shared highest similarity with Octopus tankahkeei. Through the prediction of the secondary and tertiary structure, the motor domain of KIFC1 was conserved at the C-terminal, having putative ATP-binding and microtubule-binding motifs, while the N-terminal was more specific to bind various cargoes for cellular events. The stalk domain connecting between the C-terminal and N-terminal, determined the direction of movement. According to RT-PCR results, the kifc1 gene is not tissue-specific, commonly detected in different tissues, for example, testis, liver, stomach, muscle, caecum and gills. Through an in situ hybridization method, the expression pattern of KIFC1 protein mimics in the spermatogenesis of S. maindroni. During the primary stage of the spermatogenesis, the kifc1 mRNA signal was barely detectable. At the early spermatids, the signal started to be present. With the elongation of spermatids, the signals increased substantially. It peaked and gathered around the acrosome area when the spermatids began to transform to spindle shape. As the spermatids developed into mature sperm, the signal vanished. In summary, the expression of kfic1 at specific stages during spermiogenesis and its distribution shed light on the potential functions of this motor in major cytological transformations. The KIFC1 homologue may provide a direct shaping force to the nucleus or influence the shaping process through indirect regulation.
    Full-text · Article · Sep 2013 · Gene
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