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Article ; Online: The Enigma of Centriole Loss in the 1182-4 Cell Line.

Debec, Alain / Loppin, Benjamin / Zheng, Chunfeng / Liu, Xiuwen / Megraw, Timothy L

2020 Volume 9, Issue 5

Abstract: ... ...

Abstract	The
MeSH term(s)	Animals ; Cell Line ; Centrioles/metabolism ; Drosophila Proteins/genetics ; Drosophila Proteins/metabolism ; Drosophila melanogaster/cytology ; Genome, Insect ; Models, Biological
Chemical Substances	Drosophila Proteins ; mh protein, Drosophila
Language	English
Publishing date	2020-05-23
Publishing country	Switzerland
Document type	Journal Article ; Research Support, N.I.H., Extramural ; Review
ZDB-ID	2661518-6
ISSN	2073-4409 ; 2073-4409
ISSN (online)	2073-4409
ISSN	2073-4409
DOI	10.3390/cells9051300
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Article ; Online: Basal stem cell progeny establish their apical surface in a junctional niche during turnover of an adult barrier epithelium.

Galenza, Anthony / Moreno-Roman, Paola / Su, Yu-Han / Acosta-Alvarez, Lehi / Debec, Alain / Guichet, Antoine / Knapp, Jon-Michael / Kizilyaprak, Caroline / Humbel, Bruno M / Kolotuev, Irina / O'Brien, Lucy Erin

Nature cell biology

2023 Volume 25, Issue 5, Page(s) 658–671

Abstract: Barrier epithelial organs face the constant challenge of sealing the interior body from the external environment while simultaneously replacing the cells that contact this environment. New replacement cells-the progeny of basal stem cells-are born ... ...

Abstract	Barrier epithelial organs face the constant challenge of sealing the interior body from the external environment while simultaneously replacing the cells that contact this environment. New replacement cells-the progeny of basal stem cells-are born without barrier-forming structures such as a specialized apical membrane and occluding junctions. Here, we investigate how new progeny acquire barrier structures as they integrate into the intestinal epithelium of adult Drosophila. We find they gestate their future apical membrane in a sublumenal niche created by a transitional occluding junction that envelops the differentiating cell and enables it to form a deep, microvilli-lined apical pit. The transitional junction seals the pit from the intestinal lumen until differentiation-driven, basal-to-apical remodelling of the niche opens the pit and integrates the now-mature cell into the barrier. By coordinating junctional remodelling with terminal differentiation, stem cell progeny integrate into a functional, adult epithelium without jeopardizing barrier integrity.
MeSH term(s)	Epithelium ; Cell Membrane ; Intestines ; Intestinal Mucosa/metabolism ; Stem Cells/metabolism
Language	English
Publishing date	2023-03-30
Publishing country	England
Document type	Journal Article ; Research Support, N.I.H., Extramural ; Research Support, Non-U.S. Gov't
ZDB-ID	1474722-4
ISSN	1476-4679 ; 1465-7392
ISSN (online)	1476-4679
ISSN	1465-7392
DOI	10.1038/s41556-023-01116-w
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Article ; Online: Methods to Establish Drosophila Cell Lines.

Debec, Alain / Megraw, Timothy L / Guichet, Antoine

Methods in molecular biology (Clifton, N.J.)

2016 Volume 1478, Page(s) 333–351

Abstract: Hundreds of Drosophila cell lines have been established in the labs of many researchers over the last decades and have been important tools for research. Although these cells often deviate from normal cell physiology and genetic composition, such systems ...

Abstract	Hundreds of Drosophila cell lines have been established in the labs of many researchers over the last decades and have been important tools for research. Although these cells often deviate from normal cell physiology and genetic composition, such systems nonetheless are powerful models for biochemical, cell biological, and genetics studies that are experimentally difficult in vivo. While published descriptions of cell line generation are available in the literature, how to generate new Drosophila cell lines can be challenging for beginners. Here, we describe a detailed, simple protocol to establish new Drosophila cell lines.
MeSH term(s)	Animals ; Cell Culture Techniques ; Cell Line ; Drosophila melanogaster/cytology ; Drosophila melanogaster/physiology ; Embryo, Nonmammalian/cytology ; Embryo, Nonmammalian/physiology ; Founder Effect ; Primary Cell Culture
Language	English
Publishing date	2016-09-29
Publishing country	United States
Document type	Journal Article ; Review
ISSN	1940-6029
ISSN (online)	1940-6029
DOI	10.1007/978-1-4939-6371-3_21
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Article ; Online: The Enigma of Centriole Loss in the 1182-4 Cell Line

Alain Debec / Benjamin Loppin / Chunfeng Zheng / Xiuwen Liu / Timothy L. Megraw

Cells, Vol 9, Iss 1300, p

2020 Volume 1300

Abstract: The Drosophila melanogaster cell line 1182-4, which constitutively lacks centrioles, was established many years ago from haploid embryos laid by females homozygous for the maternal haploid (mh) mutation. This was the first clear example of animal cells ... ...

Abstract	The Drosophila melanogaster cell line 1182-4, which constitutively lacks centrioles, was established many years ago from haploid embryos laid by females homozygous for the maternal haploid (mh) mutation. This was the first clear example of animal cells regularly dividing in the absence of this organelle. However, the cause of the acentriolar nature of the 1182-4 cell line remained unclear and could not be clearly assigned to a particular genetic event. Here, we detail historically the longstanding mystery of the lack of centrioles in this Drosophila cell line. Recent advances, such as the characterization of the mh gene and the genomic analysis of 1182-4 cells, allow now a better understanding of the physiology of these cells. By combining these new data, we propose three reasonable hypotheses of the genesis of this remarkable phenotype.
Keywords	centriole ; centrosome ; maternal haploid ; Drosophila melanogaster ; cell line ; haploid cells ; Biology (General) ; QH301-705.5
Language	English
Publishing date	2020-05-01T00:00:00Z
Publisher	MDPI AG
Document type	Article ; Online
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Article ; Online: Mechanisms of Horizontal Cell-to-Cell Transfer of Wolbachia spp. in Drosophila melanogaster.

White, Pamela M / Pietri, Jose E / Debec, Alain / Russell, Shelbi / Patel, Bhavin / Sullivan, William

Applied and environmental microbiology

2017 Volume 83, Issue 7

Abstract: ... ...

Abstract	Wolbachia
MeSH term(s)	Animals ; Clathrin/metabolism ; Drosophila melanogaster/cytology ; Drosophila melanogaster/microbiology ; Drosophila melanogaster/physiology ; Dynamins/metabolism ; Germ Cells/microbiology ; In Situ Hybridization, Fluorescence ; Wolbachia/cytology ; Wolbachia/physiology
Chemical Substances	Clathrin ; Dynamins (EC 3.6.5.5)
Language	English
Publishing date	2017-04-01
Publishing country	United States
Document type	Journal Article
ZDB-ID	223011-2
ISSN	1098-5336 ; 0099-2240
ISSN (online)	1098-5336
ISSN	0099-2240
DOI	10.1128/AEM.03425-16
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Article ; Online: Reliance of

White, Pamela M / Serbus, Laura R / Debec, Alain / Codina, Adan / Bray, Walter / Guichet, Antoine / Lokey, R Scott / Sullivan, William

Genetics

2017 Volume 205, Issue 4, Page(s) 1473–1488

Abstract: ... ...

Abstract	Wolbachia
MeSH term(s)	Animals ; Cell Line ; Drosophila/genetics ; Drosophila/metabolism ; Drosophila/microbiology ; Drosophila Proteins/genetics ; Drosophila Proteins/metabolism ; Endoplasmic Reticulum-Associated Degradation ; Genome, Insect ; Host-Pathogen Interactions/genetics ; Lipid Metabolism ; Mitochondria/metabolism ; Proteolysis ; RNA Interference ; Wolbachia/pathogenicity
Chemical Substances	Drosophila Proteins
Language	English
Publishing date	2017-04
Publishing country	United States
Document type	Journal Article
ZDB-ID	2167-2
ISSN	1943-2631 ; 0016-6731
ISSN (online)	1943-2631
ISSN	0016-6731
DOI	10.1534/genetics.116.198903
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Article ; Online: Centrioles: active players or passengers during mitosis?

Debec, Alain / Sullivan, William / Bettencourt-Dias, Monica

Cellular and molecular life sciences : CMLS

2010 Volume 67, Issue 13, Page(s) 2173–2194

Abstract: Centrioles are cylinders made of nine microtubule (MT) triplets present in many eukaryotes. Early studies, where centrosomes were seen at the poles of the mitotic spindle led to their coining as "the organ for cell division". However, a variety of ... ...

Abstract	Centrioles are cylinders made of nine microtubule (MT) triplets present in many eukaryotes. Early studies, where centrosomes were seen at the poles of the mitotic spindle led to their coining as "the organ for cell division". However, a variety of subsequent observational and functional studies showed that centrosomes might not always be essential for mitosis. Here we review the arguments in this debate. We describe the centriole structure and its distribution in the eukaryotic tree of life and clarify its role in the organization of the centrosome and cilia, with an historical perspective. An important aspect of the debate addressed in this review is how centrioles are inherited and the role of the spindle in this process. In particular, germline inheritance of centrosomes, such as their de novo formation in parthenogenetic species, poses many interesting questions. We finish by discussing the most likely functions of centrioles and laying out new research avenues.
MeSH term(s)	Animals ; Cell Division ; Cell Line ; Centrioles/physiology ; Centrosome/physiology ; Cilia/classification ; Cilia/metabolism ; Drosophila ; Microtubules/metabolism ; Mitosis ; Parthenogenesis ; Spindle Apparatus/physiology
Language	English
Publishing date	2010-03-19
Publishing country	Switzerland
Document type	Journal Article ; Review
ZDB-ID	1358415-7
ISSN	1420-9071 ; 1420-682X
ISSN (online)	1420-9071
ISSN	1420-682X
DOI	10.1007/s00018-010-0323-9
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Article: Centrioles: active players or passengers during mitosis

Debec, Alain / Sullivan, William / Bettencourt-Dias, Monica

Cellular and molecular life sciences CMLS. 2010 July, v. 67, no. 13

2010

Abstract: Centrioles are cylinders made of nine microtubule (MT) triplets present in many eukaryotes. Early studies, where centrosomes were seen at the poles of the mitotic spindle led to their coining as “the organ for cell division”. However, a variety of ... ...

Abstract	Centrioles are cylinders made of nine microtubule (MT) triplets present in many eukaryotes. Early studies, where centrosomes were seen at the poles of the mitotic spindle led to their coining as “the organ for cell division”. However, a variety of subsequent observational and functional studies showed that centrosomes might not always be essential for mitosis. Here we review the arguments in this debate. We describe the centriole structure and its distribution in the eukaryotic tree of life and clarify its role in the organization of the centrosome and cilia, with an historical perspective. An important aspect of the debate addressed in this review is how centrioles are inherited and the role of the spindle in this process. In particular, germline inheritance of centrosomes, such as their de novo formation in parthenogenetic species, poses many interesting questions. We finish by discussing the most likely functions of centrioles and laying out new research avenues.
Keywords	cilia ; mitosis ; parthenogenesis
Language	English
Dates of publication	2010-07
Size	p. 2173-2194.
Publisher	SP Birkhäuser Verlag Basel
Publishing place	Basel
Document type	Article
ZDB-ID	1358415-7
ISSN	1420-9071 ; 1420-682X
ISSN (online)	1420-9071
ISSN	1420-682X
DOI	10.1007/s00018-010-0323-9
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Article ; Online: Microtubule cytoskeleton remodeling by acentriolar microtubule-organizing centers at the entry and exit from mitosis in Drosophila somatic cells.

Moutinho-Pereira, Sara / Debec, Alain / Maiato, Helder

Molecular biology of the cell

2009 Volume 20, Issue 11, Page(s) 2796–2808

Abstract: Cytoskeleton microtubules undergo a reversible metamorphosis as cells enter and exit mitosis to build a transient mitotic spindle required for chromosome segregation. Centrosomes play a dominant but dispensable role in microtubule (MT) organization ... ...

Abstract	Cytoskeleton microtubules undergo a reversible metamorphosis as cells enter and exit mitosis to build a transient mitotic spindle required for chromosome segregation. Centrosomes play a dominant but dispensable role in microtubule (MT) organization throughout the animal cell cycle, supporting the existence of concurrent mechanisms that remain unclear. Here we investigated MT organization at the entry and exit from mitosis, after perturbation of centriole function in Drosophila S2 cells. We found that several MTs originate from acentriolar microtubule-organizing centers (aMTOCs) that contain gamma-tubulin and require Centrosomin (Cnn) for normal architecture and function. During spindle assembly, aMTOCs associated with peripheral MTs are recruited to acentriolar spindle poles by an Ncd/dynein-dependent clustering mechanism to form rudimentary aster-like structures. At anaphase onset, down-regulation of CDK1 triggers massive formation of cytoplasmic MTs de novo, many of which nucleated directly from aMTOCs. CDK1 down-regulation at anaphase coordinates the activity of Msps/XMAP215 and the kinesin-13 KLP10A to favor net MT growth and stability from aMTOCs. Finally, we show that microtubule nucleation from aMTOCs also occurs in cells containing centrosomes. Our data reveal a new form of cell cycle-regulated MTOCs that contribute for MT cytoskeleton remodeling during mitotic spindle assembly/disassembly in animal somatic cells, independently of centrioles.
MeSH term(s)	Anaphase/physiology ; Animals ; CDC2 Protein Kinase/genetics ; CDC2 Protein Kinase/metabolism ; Cell Line ; Centrioles/metabolism ; Cytoskeleton/metabolism ; Drosophila Proteins/genetics ; Drosophila Proteins/metabolism ; Drosophila melanogaster/cytology ; Drosophila melanogaster/metabolism ; Drosophila melanogaster/ultrastructure ; Dyneins/genetics ; Dyneins/metabolism ; Green Fluorescent Proteins/genetics ; Green Fluorescent Proteins/metabolism ; Homeodomain Proteins/genetics ; Homeodomain Proteins/metabolism ; Kinesin/genetics ; Kinesin/metabolism ; Microscopy, Electron, Transmission ; Microscopy, Fluorescence ; Microtubule-Organizing Center/metabolism ; Microtubules/metabolism ; Mitosis/physiology ; Models, Biological ; RNA Interference ; Spindle Apparatus/physiology ; Tubulin/genetics ; Tubulin/metabolism
Chemical Substances	Drosophila Proteins ; Homeodomain Proteins ; Tubulin ; cnn protein, Drosophila ; ncd protein, Drosophila ; Green Fluorescent Proteins (147336-22-9) ; CDC2 Protein Kinase (EC 2.7.11.22) ; Dyneins (EC 3.6.4.2) ; Kinesin (EC 3.6.4.4)
Language	English
Publishing date	2009-04-15
Publishing country	United States
Document type	Journal Article ; Research Support, Non-U.S. Gov't
ZDB-ID	1098979-1
ISSN	1939-4586 ; 1059-1524
ISSN (online)	1939-4586
ISSN	1059-1524
DOI	10.1091/mbc.E09-01-0011
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Article ; Online: Plk1/Polo Phosphorylates Sas-4 at the Onset of Mitosis for an Efficient Recruitment of Pericentriolar Material to Centrosomes.

Ramani, Anand / Mariappan, Aruljothi / Gottardo, Marco / Mandad, Sunit / Urlaub, Henning / Avidor-Reiss, Tomer / Riparbelli, Maria / Callaini, Giuliano / Debec, Alain / Feederle, Regina / Gopalakrishnan, Jay

Cell reports

2018 Volume 25, Issue 13, Page(s) 3618–3630.e6

Abstract: Centrosomes are the major microtubule-organizing centers, consisting of centrioles surrounded by a pericentriolar material (PCM). Centrosomal PCM is spatiotemporally regulated to be minimal during interphase and expands as cells enter mitosis. It is ... ...

Abstract	Centrosomes are the major microtubule-organizing centers, consisting of centrioles surrounded by a pericentriolar material (PCM). Centrosomal PCM is spatiotemporally regulated to be minimal during interphase and expands as cells enter mitosis. It is unclear how PCM expansion is initiated at the onset of mitosis. Here, we identify that, in Drosophila, Plk1/Polo kinase phosphorylates the conserved centrosomal protein Sas-4 in vitro. This phosphorylation appears to occur at the onset of mitosis, enabling Sas-4's localization to expand outward from meiotic and mitotic centrosomes. The Plk1/Polo kinase site of Sas-4 is then required for an efficient recruitment of Cnn and γ-tubulin, bona fide PCM proteins that are essential for PCM expansion and centrosome maturation. Point mutations at Plk1/Polo sites of Sas-4 affect neither centrosome structure nor centriole duplication but specifically reduce the affinity to bind Cnn and γ-tubulin. These observations identify Plk1/Polo kinase regulation of Sas-4 as essential for efficient PCM expansion.
MeSH term(s)	Amino Acid Sequence ; Animals ; Brain/cytology ; Centrioles/metabolism ; Centrosome/metabolism ; Drosophila Proteins/chemistry ; Drosophila Proteins/metabolism ; Drosophila melanogaster/cytology ; Drosophila melanogaster/embryology ; Drosophila melanogaster/metabolism ; Embryo, Nonmammalian/cytology ; Embryo, Nonmammalian/metabolism ; Larva/cytology ; Male ; Meiosis ; Mitosis ; Phosphorylation ; Protein Binding ; Protein Processing, Post-Translational ; Protein-Serine-Threonine Kinases/metabolism ; Spermatocytes/cytology ; Spermatocytes/metabolism
Chemical Substances	Drosophila Proteins ; Sas-4 protein, Drosophila ; polo protein, Drosophila (EC 2.7.11.-) ; Protein-Serine-Threonine Kinases (EC 2.7.11.1)
Language	English
Publishing date	2018-12-27
Publishing country	United States
Document type	Journal Article ; Research Support, Non-U.S. Gov't
ZDB-ID	2649101-1
ISSN	2211-1247 ; 2211-1247
ISSN (online)	2211-1247
ISSN	2211-1247
DOI	10.1016/j.celrep.2018.11.102
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