Roles of end‐binding 1 protein and gamma‐tubulin small complex in cytokinesis and flagella formation of Giardia lamblia
Abstract Giardia lamblia is a unicellular organism with two nuclei, a median body, eight flagella, and an adhesive disk. γ‐Tubulin is a microtubule (MT)‐nucleating protein that functions in the γ‐tubulin small complex (γ‐TuSC) in budding yeast. In this study, G. lamblia γ‐tubulin (Glγ‐tubulin) was f...
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| Format: | Article |
| Language: | English |
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Wiley
2019-06-01
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| Series: | MicrobiologyOpen |
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| Online Access: | https://doi.org/10.1002/mbo3.748 |
| _version_ | 1819241067948539904 |
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| author | Juri Kim Soon‐Jung Park |
| author_facet | Juri Kim Soon‐Jung Park |
| author_sort | Juri Kim |
| collection | DOAJ |
| description | Abstract Giardia lamblia is a unicellular organism with two nuclei, a median body, eight flagella, and an adhesive disk. γ‐Tubulin is a microtubule (MT)‐nucleating protein that functions in the γ‐tubulin small complex (γ‐TuSC) in budding yeast. In this study, G. lamblia γ‐tubulin (Glγ‐tubulin) was found to bind to another MT‐binding protein, namely G. lamblia end‐binding protein 1 (GlEB1), via both in vivo and in vitro assays. Hemagglutinin (HA)‐tagged Glγ‐tubulin localized to the basal bodies, axonemes, and median bodies of G. lamblia trophozoites. The knockdown of Glγ‐tubulin expression using an anti‐Glγ‐tubulin morpholino resulted in a decreased growth rate and an increased failed cytokinesis cells of Giardia. The formation of median bodies was affected, and the central pair of MTs in flagella was frequently missing in the Giardia treated with an anti‐Glγ‐tubulin morpholino. G. lamblia γ‐tubulin complex protein 2 (GlGCP2) and GlGCP3, which are putative components of γ‐TuSC, were co‐immunoprecipitated with HA‐tagged Glγ‐tubulin in Giardia extracts. The knockdown of GlGCP2 and GlGCP3 expression also resulted in decreased formation of both the median body and flagella MTs. Knockdown of Glγ‐tubulin, GlGCP2, and GlGCP3 expression affected localization of GlEB1 in G. lamblia. In addition, decreased level of GlEB1 caused reduced formation of median body and the central pair of flagella MTs. These results indicated that Glγ‐tubulin plays a role in MT nucleation for median body formation and flagella biogenesis as a component of Glγ‐TuSC in Giardia and GlEB1 may be involved in this process. |
| first_indexed | 2024-12-23T14:18:01Z |
| format | Article |
| id | doaj.art-5767a6fe6207408ca7c46a6a4e45ec9f |
| institution | Directory Open Access Journal |
| issn | 2045-8827 |
| language | English |
| last_indexed | 2024-12-23T14:18:01Z |
| publishDate | 2019-06-01 |
| publisher | Wiley |
| record_format | Article |
| series | MicrobiologyOpen |
| spelling | doaj.art-5767a6fe6207408ca7c46a6a4e45ec9f2022-12-21T17:43:52ZengWileyMicrobiologyOpen2045-88272019-06-0186n/an/a10.1002/mbo3.748Roles of end‐binding 1 protein and gamma‐tubulin small complex in cytokinesis and flagella formation of Giardia lambliaJuri Kim0Soon‐Jung Park1Department of Environmental Medical Biology and Institute of Tropical Medicine, Brain Korea 21 PLUS Project for Medical Science Yonsei University College of Medicine Seoul South KoreaDepartment of Environmental Medical Biology and Institute of Tropical Medicine, Brain Korea 21 PLUS Project for Medical Science Yonsei University College of Medicine Seoul South KoreaAbstract Giardia lamblia is a unicellular organism with two nuclei, a median body, eight flagella, and an adhesive disk. γ‐Tubulin is a microtubule (MT)‐nucleating protein that functions in the γ‐tubulin small complex (γ‐TuSC) in budding yeast. In this study, G. lamblia γ‐tubulin (Glγ‐tubulin) was found to bind to another MT‐binding protein, namely G. lamblia end‐binding protein 1 (GlEB1), via both in vivo and in vitro assays. Hemagglutinin (HA)‐tagged Glγ‐tubulin localized to the basal bodies, axonemes, and median bodies of G. lamblia trophozoites. The knockdown of Glγ‐tubulin expression using an anti‐Glγ‐tubulin morpholino resulted in a decreased growth rate and an increased failed cytokinesis cells of Giardia. The formation of median bodies was affected, and the central pair of MTs in flagella was frequently missing in the Giardia treated with an anti‐Glγ‐tubulin morpholino. G. lamblia γ‐tubulin complex protein 2 (GlGCP2) and GlGCP3, which are putative components of γ‐TuSC, were co‐immunoprecipitated with HA‐tagged Glγ‐tubulin in Giardia extracts. The knockdown of GlGCP2 and GlGCP3 expression also resulted in decreased formation of both the median body and flagella MTs. Knockdown of Glγ‐tubulin, GlGCP2, and GlGCP3 expression affected localization of GlEB1 in G. lamblia. In addition, decreased level of GlEB1 caused reduced formation of median body and the central pair of flagella MTs. These results indicated that Glγ‐tubulin plays a role in MT nucleation for median body formation and flagella biogenesis as a component of Glγ‐TuSC in Giardia and GlEB1 may be involved in this process.https://doi.org/10.1002/mbo3.748EB1flagellaGiardia lambliamedian bodyγ‐tubulin small complex |
| spellingShingle | Juri Kim Soon‐Jung Park Roles of end‐binding 1 protein and gamma‐tubulin small complex in cytokinesis and flagella formation of Giardia lamblia MicrobiologyOpen EB1 flagella Giardia lamblia median body γ‐tubulin small complex |
| title | Roles of end‐binding 1 protein and gamma‐tubulin small complex in cytokinesis and flagella formation of Giardia lamblia |
| title_full | Roles of end‐binding 1 protein and gamma‐tubulin small complex in cytokinesis and flagella formation of Giardia lamblia |
| title_fullStr | Roles of end‐binding 1 protein and gamma‐tubulin small complex in cytokinesis and flagella formation of Giardia lamblia |
| title_full_unstemmed | Roles of end‐binding 1 protein and gamma‐tubulin small complex in cytokinesis and flagella formation of Giardia lamblia |
| title_short | Roles of end‐binding 1 protein and gamma‐tubulin small complex in cytokinesis and flagella formation of Giardia lamblia |
| title_sort | roles of end binding 1 protein and gamma tubulin small complex in cytokinesis and flagella formation of giardia lamblia |
| topic | EB1 flagella Giardia lamblia median body γ‐tubulin small complex |
| url | https://doi.org/10.1002/mbo3.748 |
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