The Wolbachia WalE1 effector alters Drosophila endocytosis.
The most common intracellular bacterial infection is Wolbachia pipientis, a microbe that manipulates host reproduction and is used in control of insect vectors. Phenotypes induced by Wolbachia have been studied for decades and range from sperm-egg incompatibility to male killing. How Wolbachia alters host biology is less well understood. Previously, we characterized the first Wolbachia effector-WalE1, which encodes an alpha-synuclein domain at the N terminus. Purified WalE1 sediments with and bundles actin and when heterologously expressed in flies, increases Wolbachia titer in the developing oocyte. In this work, we first identify the native expression of WalE1 by Wolbachia infecting both fly cells and whole animals. WalE1 appears as aggregates in the host cell cytosol. We next show that WalE1 co-immunoprecipitates with the host protein Past1, although might not directly interact with it, and that WalE1 manipulates host endocytosis. Yeast expressing WalE1 show deficiency in uptake of FM4-64 dye, and flies harboring mutations in Past1 or overexpressing WalE1 are sensitive to AgNO3, a hallmark of endocytosis defects. We also show that flies expressing WalE1 suffer from endocytosis defects in larval nephrocytes. Finally, we also show that Past1 null flies harbor more Wolbachia overall and in late egg chambers. Our results identify interactions between Wolbachia and a host protein involved in endocytosis and point to yet another important host cell process impinged upon by Wolbachia's WalE1 effector.
Pubmed ID: 38547310 RIS Download
Research resources used in this publication
- car[1] (RRID:BDSC_19) (organism)
- y[1] w[*]; P{w[+mC]=Act5C-GAL4}17bFO1/TM6B, Tb[1] (RRID:BDSC_3954) (organism)
- y[1] w[*]; P{w[+mC]=Act5C-GAL4}25FO1/CyO, y[+] (RRID:BDSC_4414) (organism)
- w[1118]; P{w[+mC]=UAS-GFP.nls}14 (RRID:BDSC_4775) (organism)
- w[*]; sna[Sco]/CyO; Past1[60-4]/TM6B, Tb[1] (RRID:BDSC_79617) (organism)
- w[*]; sna[Sco]/CyO; Past1[110-1]/TM6B, Tb[1] (RRID:BDSC_79616) (organism)
- w[1118]; P{y[+t7.7] w[+mC]=GMR88D05-GAL4}attP2 (RRID:BDSC_48396) (organism)
- TM3, Sb[1]/TM6B, Tb[1] Dr[Mio] (RRID:BDSC_3644) (organism)
- Df(3R)kar-Sz29/TM3, Sb[1] Ser[1] (RRID:BDSC_6164) (organism)
- w[1118]; Dr[Mio]/TM3, P{w[+mC]=GAL4-twi.G}2.3, P{UAS-2xEGFP}AH2.3, Sb[1] Ser[1] (RRID:BDSC_6663) (organism)
- w[1118]; Df(3R)BSC486/TM6C, Sb[1] cu[1] (RRID:BDSC_24990) (organism)
- w[1] (RRID:BDSC_145) (organism)
- Addgene (RRID:SCR_002037) (data or information resource)
Additional research tools detected in this publication
Antibodies used in this publication
None foundAssociated grants
- Agency: NIH HHS, United States
Id: P40 OD018537 - Agency: NIAID NIH HHS, United States
Id: R01 AI144430
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GenScript (tool)
RRID:SCR_002891
Commercial organization which provides life science services and products to researchers. They specialize in gene synthesis, peptide, protein, antibody and preclinical drug development service.
View all literature mentionsThermoFisher Scientific EVOS M5000 Imaging System (tool)
RRID:SCR_023650
Microsoft system includes integrated digital inverted benchtop microscope for four-color fluorescence, transmitted-light, and color imaging. Provides interchangeable optics with autofocus, single-click multi-channel image acquisition.3.2 Megapixels, monochrome, CMOS camera. Offers software for acquisition, analysis, and automated cell counting.
View all literature mentionscar[1] (organism)
RRID:BDSC_19
Drosophila melanogaster with name car[1] from BDSC.
View all literature mentionsy[1] w[*]; P{w[+mC]=Act5C-GAL4}17bFO1/TM6B, Tb[1] (organism)
RRID:BDSC_3954
Drosophila melanogaster with name y[1] w[*]; P{w[+mC]=Act5C-GAL4}17bFO1/TM6B, Tb[1] from BDSC.
View all literature mentionsy[1] w[*]; P{w[+mC]=Act5C-GAL4}25FO1/CyO, y[+] (organism)
RRID:BDSC_4414
Drosophila melanogaster with name y[1] w[*]; P{w[+mC]=Act5C-GAL4}25FO1/CyO, y[+] from BDSC.
View all literature mentionsw[1118]; P{w[+mC]=UAS-GFP.nls}14 (organism)
RRID:BDSC_4775
Drosophila melanogaster with name w[1118]; P{w[+mC]=UAS-GFP.nls}14 from BDSC.
View all literature mentionsw[*]; sna[Sco]/CyO; Past1[60-4]/TM6B, Tb[1] (organism)
RRID:BDSC_79617
Drosophila melanogaster with name w[*]; sna[Sco]/CyO; Past1[60-4]/TM6B, Tb[1] from BDSC.
View all literature mentionsw[*]; sna[Sco]/CyO; Past1[110-1]/TM6B, Tb[1] (organism)
RRID:BDSC_79616
Drosophila melanogaster with name w[*]; sna[Sco]/CyO; Past1[110-1]/TM6B, Tb[1] from BDSC.
View all literature mentionsw[1118]; P{y[+t7.7] w[+mC]=GMR88D05-GAL4}attP2 (organism)
RRID:BDSC_48396
Drosophila melanogaster with name w[1118]; P{y[+t7.7] w[+mC]=GMR88D05-GAL4}attP2 from BDSC.
View all literature mentionsTM3, Sb[1]/TM6B, Tb[1] Dr[Mio] (organism)
RRID:BDSC_3644
Drosophila melanogaster with name TM3, Sb[1]/TM6B, Tb[1] Dr[Mio] from BDSC.
View all literature mentionsDf(3R)kar-Sz29/TM3, Sb[1] Ser[1] (organism)
RRID:BDSC_6164
Drosophila melanogaster with name Df(3R)kar-Sz29/TM3, Sb[1] Ser[1] from BDSC.
View all literature mentionsw[1118]; Dr[Mio]/TM3, P{w[+mC]=GAL4-twi.G}2.3, P{UAS-2xEGFP}AH2.3, Sb[1] Ser[1] (organism)
RRID:BDSC_6663
Drosophila melanogaster with name w[1118]; Dr[Mio]/TM3, P{w[+mC]=GAL4-twi.G}2.3, P{UAS-2xEGFP}AH2.3, Sb[1] Ser[1] from BDSC.
View all literature mentionsw[1118]; Df(3R)BSC486/TM6C, Sb[1] cu[1] (organism)
RRID:BDSC_24990
Drosophila melanogaster with name w[1118]; Df(3R)BSC486/TM6C, Sb[1] cu[1] from BDSC.
View all literature mentionsw[1] (organism)
RRID:BDSC_145
Drosophila melanogaster with name w[1] from BDSC.
View all literature mentionsAddgene (data or information resource)
RRID:SCR_002037
Non-profit plasmid repository dedicated to helping scientists around the world share high-quality plasmids. Facilitates archiving and distributing DNA-based research reagents and associated data to scientists worldwide. Repository contains over 65,000 plasmids, including special collections on CRISPR, fluorescent proteins, and ready-to-use viral preparations. There is no cost for scientists to deposit plasmids, which saves time and money associated with shipping plasmids themselves. All plasmids are fully sequenced for validation and sequencing data is openly available. We handle the appropriate Material Transfer Agreements (MTA) with institutions, facilitating open exchange and offering intellectual property and liability protection for depositing scientists. Furthermore, we curate free educational resources for the scientific community including a blog, eBooks, video protocols, and detailed molecular biology resources.
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