Vacuolar ATPase depletion affects mitochondrial ATPase function, kinetoplast dependency, and drug sensitivity in trypanosomes
Kinetoplastid parasites cause lethal diseases in humans and animals. The kinetoplast itself contains the mitochondrial genome, comprising a huge, complex DNA network that is also an important drug target. Isometamidium, for example, is a key veterinary drug that accumulates in the kinetoplast in Afr...
Saved in:
| Published in: | Proceedings of the National Academy of Sciences - PNAS Vol. 112; no. 29; p. 9112 |
|---|---|
| Main Authors: | , , , , , |
| Format: | Journal Article |
| Language: | English |
| Published: |
United States
21.07.2015
|
| Subjects: | |
| ISSN: | 1091-6490, 1091-6490 |
| Online Access: | Get more information |
| Tags: |
Add Tag
No Tags, Be the first to tag this record!
|
| Abstract | Kinetoplastid parasites cause lethal diseases in humans and animals. The kinetoplast itself contains the mitochondrial genome, comprising a huge, complex DNA network that is also an important drug target. Isometamidium, for example, is a key veterinary drug that accumulates in the kinetoplast in African trypanosomes. Kinetoplast independence and isometamidium resistance are observed where certain mutations in the F1-γ-subunit of the two-sector F1Fo-ATP synthase allow for Fo-independent generation of a mitochondrial membrane potential. To further explore kinetoplast biology and drug resistance, we screened a genome-scale RNA interference library in African trypanosomes for isometamidium resistance mechanisms. Our screen identified 14 V-ATPase subunits and all 4 adaptin-3 subunits, implicating acidic compartment defects in resistance; V-ATPase acidifies lysosomes and related organelles, whereas adaptin-3 is responsible for trafficking among these organelles. Independent strains with depleted V-ATPase or adaptin-3 subunits were isometamidium resistant, and chemical inhibition of the V-ATPase phenocopied this effect. While drug accumulation in the kinetoplast continued after V-ATPase subunit depletion, acriflavine-induced kinetoplast loss was specifically tolerated in these cells and in cells depleted for adaptin-3 or endoplasmic reticulum membrane complex subunits, also identified in our screen. Consistent with kinetoplast dispensability, V-ATPase defective cells were oligomycin resistant, suggesting ATP synthase uncoupling and bypass of the normal Fo-A6-subunit requirement; this subunit is the only kinetoplast-encoded product ultimately required for viability in bloodstream-form trypanosomes. Thus, we describe 30 genes and 3 protein complexes associated with kinetoplast-dependent growth. Mutations affecting these genes could explain natural cases of dyskinetoplasty and multidrug resistance. Our results also reveal potentially conserved communication between the compartmentalized two-sector rotary ATPases. |
|---|---|
| AbstractList | Kinetoplastid parasites cause lethal diseases in humans and animals. The kinetoplast itself contains the mitochondrial genome, comprising a huge, complex DNA network that is also an important drug target. Isometamidium, for example, is a key veterinary drug that accumulates in the kinetoplast in African trypanosomes. Kinetoplast independence and isometamidium resistance are observed where certain mutations in the F1-γ-subunit of the two-sector F1Fo-ATP synthase allow for Fo-independent generation of a mitochondrial membrane potential. To further explore kinetoplast biology and drug resistance, we screened a genome-scale RNA interference library in African trypanosomes for isometamidium resistance mechanisms. Our screen identified 14 V-ATPase subunits and all 4 adaptin-3 subunits, implicating acidic compartment defects in resistance; V-ATPase acidifies lysosomes and related organelles, whereas adaptin-3 is responsible for trafficking among these organelles. Independent strains with depleted V-ATPase or adaptin-3 subunits were isometamidium resistant, and chemical inhibition of the V-ATPase phenocopied this effect. While drug accumulation in the kinetoplast continued after V-ATPase subunit depletion, acriflavine-induced kinetoplast loss was specifically tolerated in these cells and in cells depleted for adaptin-3 or endoplasmic reticulum membrane complex subunits, also identified in our screen. Consistent with kinetoplast dispensability, V-ATPase defective cells were oligomycin resistant, suggesting ATP synthase uncoupling and bypass of the normal Fo-A6-subunit requirement; this subunit is the only kinetoplast-encoded product ultimately required for viability in bloodstream-form trypanosomes. Thus, we describe 30 genes and 3 protein complexes associated with kinetoplast-dependent growth. Mutations affecting these genes could explain natural cases of dyskinetoplasty and multidrug resistance. Our results also reveal potentially conserved communication between the compartmentalized two-sector rotary ATPases.Kinetoplastid parasites cause lethal diseases in humans and animals. The kinetoplast itself contains the mitochondrial genome, comprising a huge, complex DNA network that is also an important drug target. Isometamidium, for example, is a key veterinary drug that accumulates in the kinetoplast in African trypanosomes. Kinetoplast independence and isometamidium resistance are observed where certain mutations in the F1-γ-subunit of the two-sector F1Fo-ATP synthase allow for Fo-independent generation of a mitochondrial membrane potential. To further explore kinetoplast biology and drug resistance, we screened a genome-scale RNA interference library in African trypanosomes for isometamidium resistance mechanisms. Our screen identified 14 V-ATPase subunits and all 4 adaptin-3 subunits, implicating acidic compartment defects in resistance; V-ATPase acidifies lysosomes and related organelles, whereas adaptin-3 is responsible for trafficking among these organelles. Independent strains with depleted V-ATPase or adaptin-3 subunits were isometamidium resistant, and chemical inhibition of the V-ATPase phenocopied this effect. While drug accumulation in the kinetoplast continued after V-ATPase subunit depletion, acriflavine-induced kinetoplast loss was specifically tolerated in these cells and in cells depleted for adaptin-3 or endoplasmic reticulum membrane complex subunits, also identified in our screen. Consistent with kinetoplast dispensability, V-ATPase defective cells were oligomycin resistant, suggesting ATP synthase uncoupling and bypass of the normal Fo-A6-subunit requirement; this subunit is the only kinetoplast-encoded product ultimately required for viability in bloodstream-form trypanosomes. Thus, we describe 30 genes and 3 protein complexes associated with kinetoplast-dependent growth. Mutations affecting these genes could explain natural cases of dyskinetoplasty and multidrug resistance. Our results also reveal potentially conserved communication between the compartmentalized two-sector rotary ATPases. Kinetoplastid parasites cause lethal diseases in humans and animals. The kinetoplast itself contains the mitochondrial genome, comprising a huge, complex DNA network that is also an important drug target. Isometamidium, for example, is a key veterinary drug that accumulates in the kinetoplast in African trypanosomes. Kinetoplast independence and isometamidium resistance are observed where certain mutations in the F1-γ-subunit of the two-sector F1Fo-ATP synthase allow for Fo-independent generation of a mitochondrial membrane potential. To further explore kinetoplast biology and drug resistance, we screened a genome-scale RNA interference library in African trypanosomes for isometamidium resistance mechanisms. Our screen identified 14 V-ATPase subunits and all 4 adaptin-3 subunits, implicating acidic compartment defects in resistance; V-ATPase acidifies lysosomes and related organelles, whereas adaptin-3 is responsible for trafficking among these organelles. Independent strains with depleted V-ATPase or adaptin-3 subunits were isometamidium resistant, and chemical inhibition of the V-ATPase phenocopied this effect. While drug accumulation in the kinetoplast continued after V-ATPase subunit depletion, acriflavine-induced kinetoplast loss was specifically tolerated in these cells and in cells depleted for adaptin-3 or endoplasmic reticulum membrane complex subunits, also identified in our screen. Consistent with kinetoplast dispensability, V-ATPase defective cells were oligomycin resistant, suggesting ATP synthase uncoupling and bypass of the normal Fo-A6-subunit requirement; this subunit is the only kinetoplast-encoded product ultimately required for viability in bloodstream-form trypanosomes. Thus, we describe 30 genes and 3 protein complexes associated with kinetoplast-dependent growth. Mutations affecting these genes could explain natural cases of dyskinetoplasty and multidrug resistance. Our results also reveal potentially conserved communication between the compartmentalized two-sector rotary ATPases. |
| Author | Wilkes, Jonathan M Hutchinson, Sebastian Horn, David Barrett, Michael P Baker, Nicola Hamilton, Graham |
| Author_xml | – sequence: 1 givenname: Nicola surname: Baker fullname: Baker, Nicola organization: Division of Biological Chemistry and Drug Discovery, College of Life Sciences, University of Dundee, Dundee DD1 5EH, United Kingdom – sequence: 2 givenname: Graham surname: Hamilton fullname: Hamilton, Graham organization: Wellcome Trust Centre of Molecular Parasitology, College of Medical, Veterinary and Life Sciences, University of Glasgow, Glasgow G12 8TA, United Kingdom – sequence: 3 givenname: Jonathan M surname: Wilkes fullname: Wilkes, Jonathan M organization: Wellcome Trust Centre of Molecular Parasitology, College of Medical, Veterinary and Life Sciences, University of Glasgow, Glasgow G12 8TA, United Kingdom – sequence: 4 givenname: Sebastian surname: Hutchinson fullname: Hutchinson, Sebastian organization: Division of Biological Chemistry and Drug Discovery, College of Life Sciences, University of Dundee, Dundee DD1 5EH, United Kingdom – sequence: 5 givenname: Michael P surname: Barrett fullname: Barrett, Michael P organization: Wellcome Trust Centre of Molecular Parasitology, College of Medical, Veterinary and Life Sciences, University of Glasgow, Glasgow G12 8TA, United Kingdom – sequence: 6 givenname: David surname: Horn fullname: Horn, David email: d.horn@dundee.ac.uk organization: Division of Biological Chemistry and Drug Discovery, College of Life Sciences, University of Dundee, Dundee DD1 5EH, United Kingdom; d.horn@dundee.ac.uk |
| BackLink | https://www.ncbi.nlm.nih.gov/pubmed/26150481$$D View this record in MEDLINE/PubMed |
| BookMark | eNpNkD1PwzAYhC1URD9gZkMeGZpiJ3Ecj1XFl1QJhsJaOfZrMCR2iB2kDvx3UtFK3HI3PHfDTdHIeQcIXVKyoIRnN62TYUEZYTkdlJ6gCSWCJkUuyOhfHqNpCB-EEMFKcobGaTF08pJO0M-rVL2vZYeXm2cZAGtoa4jWOyyNARUDbmz06t073VlZHzHTO7Wn5vjTOoi-rWWI-zI4DU7t5lg6jXXXv-EALthov23cYetw7HatdD74BsI5OjWyDnBx8Bl6ubvdrB6S9dP942q5ThRjPCZasSrLCeVCZFxxwyEzOpWKac0LRrgxTLCKm8zQohRUpcRQzmSWS5B5Qat0hq7_dtvOf_UQ4raxQUFdSwe-D1taiDIT6XDJgF4d0L5qQG_bzjay222Pl6W_60dylg |
| CitedBy_id | crossref_primary_10_1242_bio_048926 crossref_primary_10_1038_s41467_024_55538_7 crossref_primary_10_3390_cells11121977 crossref_primary_10_1016_j_mib_2016_04_019 crossref_primary_10_1038_s42003_025_08086_9 crossref_primary_10_1371_journal_ppat_1009717 crossref_primary_10_1128_AAC_02268_17 crossref_primary_10_1038_s41440_020_0450_0 crossref_primary_10_1007_s11686_022_00640_3 crossref_primary_10_1016_j_actatropica_2019_105227 crossref_primary_10_1096_fj_202001579RR crossref_primary_10_1016_j_molbiopara_2015_10_003 crossref_primary_10_3390_molecules23092331 crossref_primary_10_7554_eLife_32496 crossref_primary_10_1017_S0031182016001268 crossref_primary_10_3390_genes9070368 crossref_primary_10_1038_s41422_023_00820_4 crossref_primary_10_1371_journal_pntd_0004791 crossref_primary_10_1016_j_pt_2021_09_002 crossref_primary_10_1186_s13071_020_04148_5 crossref_primary_10_1371_journal_pntd_0006790 crossref_primary_10_1016_j_molbiopara_2024_111619 crossref_primary_10_1016_j_molmed_2019_10_009 crossref_primary_10_1007_s00253_023_12875_9 crossref_primary_10_1371_journal_ppat_1006855 crossref_primary_10_1186_s13071_020_04068_4 crossref_primary_10_1371_journal_pgen_1009603 crossref_primary_10_1371_journal_pone_0146511 crossref_primary_10_1128_AAC_00235_18 crossref_primary_10_1371_journal_ppat_1006477 crossref_primary_10_1074_jbc_M116_714972 crossref_primary_10_1002_mc_23741 crossref_primary_10_1371_journal_pntd_0004556 |
| ContentType | Journal Article |
| DBID | CGR CUY CVF ECM EIF NPM 7X8 |
| DOI | 10.1073/pnas.1505411112 |
| DatabaseName | Medline MEDLINE MEDLINE (Ovid) MEDLINE MEDLINE PubMed MEDLINE - Academic |
| DatabaseTitle | MEDLINE Medline Complete MEDLINE with Full Text PubMed MEDLINE (Ovid) MEDLINE - Academic |
| DatabaseTitleList | MEDLINE - Academic MEDLINE |
| Database_xml | – sequence: 1 dbid: NPM name: PubMed url: http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed sourceTypes: Index Database – sequence: 2 dbid: 7X8 name: MEDLINE - Academic url: https://search.proquest.com/medline sourceTypes: Aggregation Database |
| DeliveryMethod | no_fulltext_linktorsrc |
| Discipline | Sciences (General) |
| EISSN | 1091-6490 |
| ExternalDocumentID | 26150481 |
| Genre | Research Support, Non-U.S. Gov't Journal Article |
| GrantInformation_xml | – fundername: Wellcome Trust grantid: 100320/Z/12/Z – fundername: Wellcome Trust grantid: 083481/Z/07/Z – fundername: Wellcome Trust grantid: 100320 – fundername: Wellcome Trust grantid: 093010/Z/10/Z – fundername: Wellcome Trust grantid: 085349 |
| GroupedDBID | --- -DZ -~X .55 0R~ 123 29P 2AX 2FS 2WC 4.4 53G 5RE 5VS 85S AACGO AAFWJ AANCE ABBHK ABOCM ABPLY ABPPZ ABTLG ABXSQ ABZEH ACGOD ACHIC ACIWK ACNCT ACPRK ADQXQ ADULT AENEX AEUPB AEXZC AFFNX AFOSN AFRAH ALMA_UNASSIGNED_HOLDINGS AQVQM BKOMP CGR CS3 CUY CVF D0L DCCCD DIK DU5 E3Z EBS ECM EIF EJD F5P FRP GX1 H13 HH5 HYE IPSME JAAYA JBMMH JENOY JHFFW JKQEH JLS JLXEF JPM JSG JST KQ8 L7B LU7 N9A NPM N~3 O9- OK1 PNE PQQKQ R.V RHI RNA RNS RPM RXW SA0 SJN TAE TN5 UKR W8F WH7 WOQ WOW X7M XSW Y6R YBH YKV YSK ZCA ~02 ~KM 7X8 |
| ID | FETCH-LOGICAL-c557t-dc5b340179937c7f7e3fd2ac5dd76507ff595b7f3f16891c20f175a34aea461b2 |
| IEDL.DBID | 7X8 |
| ISICitedReferencesCount | 41 |
| ISICitedReferencesURI | http://www.webofscience.com/api/gateway?GWVersion=2&SrcApp=Summon&SrcAuth=ProQuest&DestLinkType=CitingArticles&DestApp=WOS_CPL&KeyUT=000358225100084&url=https%3A%2F%2Fcvtisr.summon.serialssolutions.com%2F%23%21%2Fsearch%3Fho%3Df%26include.ft.matches%3Dt%26l%3Dnull%26q%3D |
| ISSN | 1091-6490 |
| IngestDate | Fri Sep 05 06:19:28 EDT 2025 Sat May 31 02:12:17 EDT 2025 |
| IsDoiOpenAccess | false |
| IsOpenAccess | true |
| IsPeerReviewed | true |
| IsScholarly | true |
| Issue | 29 |
| Keywords | samorin brucei nagana mitochondrion petite |
| Language | English |
| LinkModel | DirectLink |
| MergedId | FETCHMERGED-LOGICAL-c557t-dc5b340179937c7f7e3fd2ac5dd76507ff595b7f3f16891c20f175a34aea461b2 |
| Notes | ObjectType-Article-1 SourceType-Scholarly Journals-1 ObjectType-Feature-2 content type line 23 |
| OpenAccessLink | https://www.ncbi.nlm.nih.gov/pmc/articles/4517229 |
| PMID | 26150481 |
| PQID | 1698392048 |
| PQPubID | 23479 |
| ParticipantIDs | proquest_miscellaneous_1698392048 pubmed_primary_26150481 |
| PublicationCentury | 2000 |
| PublicationDate | 2015-07-21 |
| PublicationDateYYYYMMDD | 2015-07-21 |
| PublicationDate_xml | – month: 07 year: 2015 text: 2015-07-21 day: 21 |
| PublicationDecade | 2010 |
| PublicationPlace | United States |
| PublicationPlace_xml | – name: United States |
| PublicationTitle | Proceedings of the National Academy of Sciences - PNAS |
| PublicationTitleAlternate | Proc Natl Acad Sci U S A |
| PublicationYear | 2015 |
| SSID | ssj0009580 |
| Score | 2.3733425 |
| Snippet | Kinetoplastid parasites cause lethal diseases in humans and animals. The kinetoplast itself contains the mitochondrial genome, comprising a huge, complex DNA... |
| SourceID | proquest pubmed |
| SourceType | Aggregation Database Index Database |
| StartPage | 9112 |
| SubjectTerms | Acids - metabolism Animals Cell Compartmentation - drug effects DNA, Kinetoplast - metabolism Drug Resistance - drug effects Endoplasmic Reticulum - drug effects Endoplasmic Reticulum - metabolism Intracellular Membranes - drug effects Intracellular Membranes - metabolism Mitochondria - drug effects Mitochondria - enzymology Phenanthridines - chemistry Phenanthridines - pharmacology Polymerase Chain Reaction Protein Subunits - metabolism Proton-Translocating ATPases - metabolism Reproducibility of Results RNA Interference - drug effects Trypanosoma brucei brucei - drug effects Trypanosoma brucei brucei - enzymology Trypanosoma brucei brucei - genetics Vacuolar Proton-Translocating ATPases - metabolism |
| Title | Vacuolar ATPase depletion affects mitochondrial ATPase function, kinetoplast dependency, and drug sensitivity in trypanosomes |
| URI | https://www.ncbi.nlm.nih.gov/pubmed/26150481 https://www.proquest.com/docview/1698392048 |
| Volume | 112 |
| WOSCitedRecordID | wos000358225100084&url=https%3A%2F%2Fcvtisr.summon.serialssolutions.com%2F%23%21%2Fsearch%3Fho%3Df%26include.ft.matches%3Dt%26l%3Dnull%26q%3D |
| hasFullText | |
| inHoldings | 1 |
| isFullTextHit | |
| isPrint | |
| link | http://cvtisr.summon.serialssolutions.com/2.0.0/link/0/eLvHCXMwpZ3Pa9swFMfF1u6wS9d0W5t1HRr00EG8WLZkWacRSssOW8ihLbkFWT9GGJHd2Cnk0P-978U222VQ2MVgkMHI78fH0tP3EXIuU6917IpIM24j7q2NlIJbJb0TViXS7c5W3f2Q02k-n6tZt-BWd2WVfUzcBWpbGlwjH7NMYS4Hg_tW3UfYNQp3V7sWGi_JfgoogyVdcp7_Jbqbt2oEikUZV3Ev7SPTcRV0_RVgSHCMGcm_-XKXZ67f_O8bHpKDjjDppDWJAXnhwhEZdD5c04tOaPrLW_J4p80Gf23p5GYG2YxaV6EWdxmobss86AocHgJksGin_TDMhThqRH8DozZlBQTe0L6drtmOqA6W2vXmF62xPr5tUEGXgTbrLUSfsi5Xrn5Hbq-vbi6_R107hsgIIZvIGlGkHD0YkMZIL13qbaKNsFYC50nvhRKF9KlnWa6YSWIPbKJTrp3mGSuS92QvlMGdEArMKEVR5Hiql1sZKy8yk2lmmIx17Ish-dxP8QLMHfcwdHDlpl78meQhOW6_06JqdTkWCYrb85x9eMbTp-Q1oI_AVdqEfST7HpzdnZFX5qFZ1utPOzuC63T28wmeq9WN |
| linkProvider | ProQuest |
| openUrl | ctx_ver=Z39.88-2004&ctx_enc=info%3Aofi%2Fenc%3AUTF-8&rfr_id=info%3Asid%2Fsummon.serialssolutions.com&rft_val_fmt=info%3Aofi%2Ffmt%3Akev%3Amtx%3Ajournal&rft.genre=article&rft.atitle=Vacuolar+ATPase+depletion+affects+mitochondrial+ATPase+function%2C+kinetoplast+dependency%2C+and+drug+sensitivity+in+trypanosomes&rft.jtitle=Proceedings+of+the+National+Academy+of+Sciences+-+PNAS&rft.au=Baker%2C+Nicola&rft.au=Hamilton%2C+Graham&rft.au=Wilkes%2C+Jonathan+M&rft.au=Hutchinson%2C+Sebastian&rft.date=2015-07-21&rft.eissn=1091-6490&rft.volume=112&rft.issue=29&rft.spage=9112&rft_id=info:doi/10.1073%2Fpnas.1505411112&rft_id=info%3Apmid%2F26150481&rft_id=info%3Apmid%2F26150481&rft.externalDocID=26150481 |
| thumbnail_l | http://covers-cdn.summon.serialssolutions.com/index.aspx?isbn=/lc.gif&issn=1091-6490&client=summon |
| thumbnail_m | http://covers-cdn.summon.serialssolutions.com/index.aspx?isbn=/mc.gif&issn=1091-6490&client=summon |
| thumbnail_s | http://covers-cdn.summon.serialssolutions.com/index.aspx?isbn=/sc.gif&issn=1091-6490&client=summon |