The Lyme disease agent co-opts adiponectin receptor-mediated signaling in its arthropod vector
- Section of Infectious Diseases, Department of Internal Medicine, School of Medicine, Yale University, United States
- Department of Clinical Veterinary Medicine, and Key Laboratory for Zoonosis Research, Ministry of Education, College of Veterinary Medicine, Jilin University, China
- Yale Combined Program in the Biological and Biomedical Sciences, Yale University, United States
- Yale Center for Genome Analysis, Yale University, United States
- Department of Microbiology, School of Basic Medicine, Fourth Military Medical University, China
- Department of Veterinary Medicine, University of Maryland, College Park, United States
Figures
Figure 1 with 2 supplements
Silencing of ISARL significantly reduces the B. burgdorferi burden in nymphal tick guts.
(A) ISARL is significantly induced in nymphal tick guts after feeding on B. burgdorferi-infected mice. (B) qPCR assessment of ISARL transcript levels following RNAi silencing of ISARL after feeding …
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Figure 1—source data 1
ISARL is involved in B. burgdorferi colonization in nymphal tick guts but has no effect on transmission.
- https://cdn.elifesciences.org/articles/72568/elife-72568-fig1-data1-v3.xlsx
Figure 1—figure supplement 1
Protein sequence comparison of adiponectin receptors.
Multiple sequence alignment of Ixodes scapularis ISARL with the amino acid sequences of homologs identified in Homo sapiens (NP_001277482, HsAdipoR1; NP_001362293, HsAdipoR2), Mus musculus …
Figure 1—figure supplement 2
Predicted protein structure and hydrophobicity of ISARL.
Seven transmembrane (TM) domains were identified in the ISARL protein based on (A) protein structure prediction and (B) hydrophobicity analysis.
Figure 2 with 3 supplements
RNA-seq, qPCR validation, and RNAi-silencing assays revealed that phosphatidylserine synthase 1 (PTDSS1) is regulated by ISARL and is involved in B. burgdorferi colonization.
(A) Hierarchical clustering of differentially expressed genes was generated after feeding on clean mice. (B) Hierarchical clustering of differentially expressed genes was generated after feeding on B…
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Figure 2—source data 1
Source data for PTDSS1 protein relative quantification.
- https://cdn.elifesciences.org/articles/72568/elife-72568-fig2-data1-v3.pdf
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Figure 2—source data 2
Source data for PTDSS1 protein relative quantification.
- https://cdn.elifesciences.org/articles/72568/elife-72568-fig2-data2-v3.pdf
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Figure 2—source data 3
PTDSS1 is regulated by ISARL and is involved in B. burgdorferi colonization.
- https://cdn.elifesciences.org/articles/72568/elife-72568-fig2-data3-v3.xlsx
Figure 2—figure supplement 1
qPCR validation of 18 well-annotated and metabolism-related differentially expressed genes.
3HADH, 3-hydroxyacyl-CoA dehydrogenase, putative; ADSS, adenylosuccinate synthetase; GMPS, GMP synthase, putative; PDE, cAMP and cAMP-inhibited cGMP 3,5-cyclic phosphodiesterase; GP, glycogen …
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Figure 2—figure supplement 1—source data 1
Source data for qPCR validation.
- https://cdn.elifesciences.org/articles/72568/elife-72568-fig2-figsupp1-data1-v3.xlsx
Figure 2—figure supplement 2
QPCR validation of differentially expressed genes from the RNA-seq dataset.
(A) N-CAM Ig domain-containing protein (NCAM), vacuolar H+-ATPase V1 sector, subunit G (V-ATPase), and sideroflexin 1,2,3, putative (SFXN) were significantly downregulated following RNAi silencing …
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Figure 2—figure supplement 2—source data 1
Source data for qPCR validation.
- https://cdn.elifesciences.org/articles/72568/elife-72568-fig2-figsupp2-data1-v3.xlsx
Figure 2—figure supplement 3
Silencing of differentially expressed genes and effects on B. burgdorferi acquisition.
Silencing of (A) NCAM, (B) V-ATPase, (C) SFXN, (E) ADSS, (F) GMPS, and (G) ACTN has no effect on B. burgdorferi acquisition. Silencing of (D) 3HADH decreased the B. burgdorferi burden in tick gut. …
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Figure 2—figure supplement 3—source data 1
Source data for RNAi of differentially expressed genes and effects on B. burgdorferi acquisition.
- https://cdn.elifesciences.org/articles/72568/elife-72568-fig2-figsupp3-data1-v3.xlsx
Figure 3 with 1 supplement
Mammalian adiponectin regulates tick glucose metabolism.
(A) RNA-seq of injection of recombinant mouse adiponectin and GFP (control) proteins. One classic downstream gene of mammalian adiponectin receptor signaling, glucose-6-phosphatase (G6p), was …
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Figure 3—source data 1
Mammalian adiponectin regulates tick glucose metabolism but has no effect on B. burgdorferi colonization.
- https://cdn.elifesciences.org/articles/72568/elife-72568-fig3-data1-v3.xlsx
Figure 3—figure supplement 1
Mammalian adiponectin and tick glucose metabolism changes have no effect on B. burgdorferi acquisition.
(A) qPCR assessment of G6pc1 transcript level, nymphal engorgement weights, and qPCR assessment of B. burgdorferi flaB levels in guts following RNAi silencing of G6pc1 after feeding on B. burgdorferi…
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Figure 3—figure supplement 1—source data 1
Source data for effects of tick glucose metabolism on B. burgdorferi acquisition.
- https://cdn.elifesciences.org/articles/72568/elife-72568-fig3-figsupp1-data1-v3.xlsx
Figure 4 with 1 supplement
Tick complement C1q-like protein 3 (C1QL3) is involved in ISARL signaling pathways.
(A) Blastp of the tick genome with the human adiponectin C1Q domain in NCBI generated two homologs and were annotated as complement C1q-like protein 3 (C1QL3) (GenBank number: XP_002415101) and …
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Figure 4—source data 1
Source data for ISARL expression.
- https://cdn.elifesciences.org/articles/72568/elife-72568-fig4-data1-v3.pdf
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Figure 4—source data 2
Source data for C1QL3 protein purification.
- https://cdn.elifesciences.org/articles/72568/elife-72568-fig4-data2-v3.pdf
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Figure 4—source data 3
Source data for C1QL3 protein purification.
- https://cdn.elifesciences.org/articles/72568/elife-72568-fig4-data3-v3.pdf
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Figure 4—source data 4
Source data for binding of C1QL3 to ISARL.
- https://cdn.elifesciences.org/articles/72568/elife-72568-fig4-data4-v3.pdf
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Figure 4—source data 5
Source data for binding of C1QL3 to tick ISE6 cells.
- https://cdn.elifesciences.org/articles/72568/elife-72568-fig4-data5-v3.pdf
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Figure 4—source data 6
C1QL3 is involved in the ISARL signaling pathway and modulates B. burgdorferi colonization.
- https://cdn.elifesciences.org/articles/72568/elife-72568-fig4-data6-v3.xlsx
Figure 4—figure supplement 1
Alignment of human adiponectin C1Q domain and C1QL3 C1Q domain.
Tick C1QL3 C1Q domain has 64.0% similarity and 28.8% identity with human adiponectin C1Q domain. The alignment was conducted in LALIGN/PLALIGN (https://fasta.bioch.virginia.edu/fasta_www2/fasta_www.c…
Tables
Key resources table
| Reagent type (species) or resource | Designation | Source or reference | Identifiers | Additional information |
|---|---|---|---|---|
| Biological sample (Mus musculus) | C3H/HeJ | Jackson Laboratory | Stock #: 000659;RRID:IMSR_JAX:000659 | |
| Biological sample (M. musculus) | WT C57BL/6J | Jackson Laboratory | Stock #: 000664; RRID:IMSR_JAX:000664 | |
| Biological sample (M. musculus) | Adipoq-/- C57BL/6J | Jackson Laboratory | Stock #: 008195; RRID:IMSR_JAX:008195 | |
| Biological sample (Borrelia burgdorferi) | Borrelia burgdorferi strain N40 | Dr. Erol Fikrig Laboratory | ||
| Biological sample (Ixodes scapularis) | Black-legged tick | Oklahoma State University | ||
| Cell line (Homo sapiens) | Human embryonic kidney HEK293T | ATCC | #CRL-3216; RRID:CVCL_0063 | |
| Cell line (I. scapularis) | Tick ISE6 | ATCC | #CRL-11974; RRID:CVCL_Z170 | |
| Antibody | Anti-HA (rabbit monoclonal) | Cell Signaling Technology | #C29F4; RRID:AB_10693385 | IF (1:100) |
| Antibody | Anti-V5 (mouse monoclonal) | Invitrogen | #R960-25; RRID:AB_2556564 | IF (1:100) |
| Antibody | Goat anti-rabbit IgG (H + L) Highly Cross-Adsorbed Secondary Antibody, Alexa Fluor 488 | Invitrogen | #A-11034; RRID:AB_2576217 | IF (1:100) |
| Antibody | Goat anti-mouse IgG (H + L) Cross-Adsorbed Secondary Antibody, Alexa Fluor 555 | Invitrogen | #A-21422; RRID:AB_141822 | IF (1:100) |
| Antibody | HRP Anti-His tag antibody (chicken polyclonal) | Abcam | #ab3553; RRID:AB_303900 | WB (1:10,000) |
| Antibody | HRP V5-tag (mouse monoclonal) | Invitrogen | #R961-25; RRID:AB_2556565 | WB (1:1000) |
| Peptide, recombinant protein | Mouse adiponectin | MilliporeSigma | #SRP3297 | |
| Peptide, recombinant protein | Aequorea victoria green fluorescent protein (GFP) | SinoBiological | #13105-S07E | |
| Commercial assay or kit | Mouse adiponectin/Acrp30 Quantikine ELISA Kit | R&D Systems | #MRP300;RRID:AB_2832917 | |
| Software, algorithm | Prism | GraphPad | RRID:SCR_002798 |
Additional files
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Transparent reporting form
- https://cdn.elifesciences.org/articles/72568/elife-72568-transrepform1-v3.docx
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Supplementary file 1
Supplementary files in this study.
(a) Summary of differently expressed genes of comparison between ds GFP and ds ISARL injection after 96 hr feeding on clean mice. (b) Summary of differently expressed genes of comparison between ds GFP and ds ISARL injection after 96 hr feeding on B. burgdorferi-infected mice. (c) Summary of differently expressed genes of comparison between recombinant GFP and adiponectin proteins injection after 8 hr. (d) The primers used in this study.
- https://cdn.elifesciences.org/articles/72568/elife-72568-supp1-v3.docx
