The intracellular delivery of TAT-aequorin reveals calcium-mediated sensing of environmental and symbiotic signals by the arbuscular mycorrhizal fungus Gigaspora margarita.
Identifieur interne : 001861 ( Main/Corpus ); précédent : 001860; suivant : 001862The intracellular delivery of TAT-aequorin reveals calcium-mediated sensing of environmental and symbiotic signals by the arbuscular mycorrhizal fungus Gigaspora margarita.
Auteurs : Roberto Moscatiello ; Simone Sello ; Mara Novero ; Alessandro Negro ; Paola Bonfante ; Lorella NavazioSource :
- The New phytologist [ 1469-8137 ] ; 2014.
English descriptors
- KwdEn :
- Aequorin (metabolism), Calcium (metabolism), Endocytosis (drug effects), Environment (MeSH), Gene Transfer Techniques (MeSH), Glomeromycota (drug effects), Glomeromycota (physiology), Green Fluorescent Proteins (metabolism), Hyphae (drug effects), Hyphae (metabolism), Immunoblotting (MeSH), Intracellular Space (drug effects), Intracellular Space (metabolism), Lactones (pharmacology), Luminescent Measurements (MeSH), Mycorrhizae (drug effects), Mycorrhizae (physiology), Peptides (metabolism), Symbiosis (drug effects), Symbiosis (physiology), tat Gene Products, Human Immunodeficiency Virus (metabolism).
- MESH :
- chemical , metabolism : Aequorin, Calcium, Green Fluorescent Proteins, Peptides, tat Gene Products, Human Immunodeficiency Virus.
- drug effects : Endocytosis, Glomeromycota, Hyphae, Intracellular Space, Mycorrhizae, Symbiosis.
- metabolism : Hyphae, Intracellular Space.
- chemical , pharmacology : Lactones.
- physiology : Glomeromycota, Mycorrhizae, Symbiosis.
- Environment, Gene Transfer Techniques, Immunoblotting, Luminescent Measurements.
Abstract
Arbuscular mycorrhiza (AM) is an ecologically relevant symbiosis between most land plants and Glomeromycota fungi. The peculiar traits of AM fungi have so far limited traditional approaches such as genetic transformation. The aim of this work was to investigate whether the protein transduction domain of the HIV-1 transactivator of transcription (TAT) protein, previously shown to act as a potent nanocarrier for macromolecule delivery in both animal and plant cells, may translocate protein cargoes into AM fungi. We evaluated the internalization into germinated spores of Gigaspora margarita of two recombinant TAT fusion proteins consisting of either a fluorescent (GFP) or a luminescent (aequorin) reporter linked to the TAT peptide. Both TAT-fused proteins were found to enter AM fungal mycelia after a short incubation period (5-10 min). Ca2+ measurements in G. margarita mycelia pre-incubated with TAT-aequorin demonstrated the occurrence of changes in the intracellular free Ca2+ concentration in response to relevant stimuli, such as touch, cold, salinity, and strigolactones, symbiosis-related plant signals. These data indicate that the cell-penetrating properties of the TAT peptide can be used as an effective strategy for intracellularly delivering proteins of interest and shed new light on Ca2+ homeostasis and signalling in AM fungi.
DOI: 10.1111/nph.12849
PubMed: 24845011
Links to Exploration step
pubmed:24845011Le document en format XML
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Bassi 58/B, 35131, Padova, Italy.</nlm:affiliation></affiliation></author><author><name sortKey="Sello, Simone" sort="Sello, Simone" uniqKey="Sello S" first="Simone" last="Sello">Simone Sello</name></author><author><name sortKey="Novero, Mara" sort="Novero, Mara" uniqKey="Novero M" first="Mara" last="Novero">Mara Novero</name></author><author><name sortKey="Negro, Alessandro" sort="Negro, Alessandro" uniqKey="Negro A" first="Alessandro" last="Negro">Alessandro Negro</name></author><author><name sortKey="Bonfante, Paola" sort="Bonfante, Paola" uniqKey="Bonfante P" first="Paola" last="Bonfante">Paola Bonfante</name></author><author><name sortKey="Navazio, Lorella" sort="Navazio, Lorella" uniqKey="Navazio L" first="Lorella" last="Navazio">Lorella Navazio</name></author></titleStmt><publicationStmt><idno type="wicri:source">PubMed</idno><date when="2014">2014</date><idno type="RBID">pubmed:24845011</idno><idno type="pmid">24845011</idno><idno type="doi">10.1111/nph.12849</idno><idno type="wicri:Area/Main/Corpus">001861</idno><idno type="wicri:explorRef" wicri:stream="Main" wicri:step="Corpus" wicri:corpus="PubMed">001861</idno></publicationStmt><sourceDesc><biblStruct><analytic><title xml:lang="en">The intracellular delivery of TAT-aequorin reveals calcium-mediated sensing of environmental and symbiotic signals by the arbuscular mycorrhizal fungus Gigaspora margarita.</title><author><name sortKey="Moscatiello, Roberto" sort="Moscatiello, Roberto" uniqKey="Moscatiello R" first="Roberto" last="Moscatiello">Roberto Moscatiello</name><affiliation><nlm:affiliation>Department of Biology, University of Padova, Via U. Bassi 58/B, 35131, Padova, Italy.</nlm:affiliation></affiliation></author><author><name sortKey="Sello, Simone" sort="Sello, Simone" uniqKey="Sello S" first="Simone" last="Sello">Simone Sello</name></author><author><name sortKey="Novero, Mara" sort="Novero, Mara" uniqKey="Novero M" first="Mara" last="Novero">Mara Novero</name></author><author><name sortKey="Negro, Alessandro" sort="Negro, Alessandro" uniqKey="Negro A" first="Alessandro" last="Negro">Alessandro Negro</name></author><author><name sortKey="Bonfante, Paola" sort="Bonfante, Paola" uniqKey="Bonfante P" first="Paola" last="Bonfante">Paola Bonfante</name></author><author><name sortKey="Navazio, Lorella" sort="Navazio, Lorella" uniqKey="Navazio L" first="Lorella" last="Navazio">Lorella Navazio</name></author></analytic><series><title level="j">The New phytologist</title><idno type="eISSN">1469-8137</idno><imprint><date when="2014" type="published">2014</date></imprint></series></biblStruct></sourceDesc></fileDesc><profileDesc><textClass><keywords scheme="KwdEn" xml:lang="en"><term>Aequorin (metabolism)</term><term>Calcium (metabolism)</term><term>Endocytosis (drug effects)</term><term>Environment (MeSH)</term><term>Gene Transfer Techniques (MeSH)</term><term>Glomeromycota (drug effects)</term><term>Glomeromycota (physiology)</term><term>Green Fluorescent Proteins (metabolism)</term><term>Hyphae (drug effects)</term><term>Hyphae (metabolism)</term><term>Immunoblotting (MeSH)</term><term>Intracellular Space (drug effects)</term><term>Intracellular Space (metabolism)</term><term>Lactones (pharmacology)</term><term>Luminescent Measurements (MeSH)</term><term>Mycorrhizae (drug effects)</term><term>Mycorrhizae (physiology)</term><term>Peptides (metabolism)</term><term>Symbiosis (drug effects)</term><term>Symbiosis (physiology)</term><term>tat Gene Products, Human Immunodeficiency Virus (metabolism)</term></keywords><keywords scheme="MESH" type="chemical" qualifier="metabolism" xml:lang="en"><term>Aequorin</term><term>Calcium</term><term>Green Fluorescent Proteins</term><term>Peptides</term><term>tat Gene Products, Human Immunodeficiency Virus</term></keywords><keywords scheme="MESH" qualifier="drug effects" xml:lang="en"><term>Endocytosis</term><term>Glomeromycota</term><term>Hyphae</term><term>Intracellular Space</term><term>Mycorrhizae</term><term>Symbiosis</term></keywords><keywords scheme="MESH" qualifier="metabolism" xml:lang="en"><term>Hyphae</term><term>Intracellular Space</term></keywords><keywords scheme="MESH" type="chemical" qualifier="pharmacology" xml:lang="en"><term>Lactones</term></keywords><keywords scheme="MESH" qualifier="physiology" xml:lang="en"><term>Glomeromycota</term><term>Mycorrhizae</term><term>Symbiosis</term></keywords><keywords scheme="MESH" xml:lang="en"><term>Environment</term><term>Gene Transfer Techniques</term><term>Immunoblotting</term><term>Luminescent Measurements</term></keywords></textClass></profileDesc></teiHeader><front><div type="abstract" xml:lang="en">Arbuscular mycorrhiza (AM) is an ecologically relevant symbiosis between most land plants and Glomeromycota fungi. The peculiar traits of AM fungi have so far limited traditional approaches such as genetic transformation. The aim of this work was to investigate whether the protein transduction domain of the HIV-1 transactivator of transcription (TAT) protein, previously shown to act as a potent nanocarrier for macromolecule delivery in both animal and plant cells, may translocate protein cargoes into AM fungi. We evaluated the internalization into germinated spores of Gigaspora margarita of two recombinant TAT fusion proteins consisting of either a fluorescent (GFP) or a luminescent (aequorin) reporter linked to the TAT peptide. Both TAT-fused proteins were found to enter AM fungal mycelia after a short incubation period (5-10 min). Ca2+ measurements in G. margarita mycelia pre-incubated with TAT-aequorin demonstrated the occurrence of changes in the intracellular free Ca2+ concentration in response to relevant stimuli, such as touch, cold, salinity, and strigolactones, symbiosis-related plant signals. These data indicate that the cell-penetrating properties of the TAT peptide can be used as an effective strategy for intracellularly delivering proteins of interest and shed new light on Ca2+ homeostasis and signalling in AM fungi.</div></front></TEI><pubmed><MedlineCitation Status="MEDLINE" Owner="NLM"><PMID Version="1">24845011</PMID><DateCompleted><Year>2015</Year><Month>03</Month><Day>30</Day></DateCompleted><DateRevised><Year>2020</Year><Month>09</Month><Day>30</Day></DateRevised><Article PubModel="Print-Electronic"><Journal><ISSN IssnType="Electronic">1469-8137</ISSN><JournalIssue CitedMedium="Internet"><Volume>203</Volume><Issue>3</Issue><PubDate><Year>2014</Year><Month>Aug</Month></PubDate></JournalIssue><Title>The New phytologist</Title><ISOAbbreviation>New Phytol</ISOAbbreviation></Journal><ArticleTitle>The intracellular delivery of TAT-aequorin reveals calcium-mediated sensing of environmental and symbiotic signals by the arbuscular mycorrhizal fungus Gigaspora margarita.</ArticleTitle><Pagination><MedlinePgn>1012-20</MedlinePgn></Pagination><ELocationID EIdType="doi" ValidYN="Y">10.1111/nph.12849</ELocationID><Abstract><AbstractText>Arbuscular mycorrhiza (AM) is an ecologically relevant symbiosis between most land plants and Glomeromycota fungi. The peculiar traits of AM fungi have so far limited traditional approaches such as genetic transformation. The aim of this work was to investigate whether the protein transduction domain of the HIV-1 transactivator of transcription (TAT) protein, previously shown to act as a potent nanocarrier for macromolecule delivery in both animal and plant cells, may translocate protein cargoes into AM fungi. We evaluated the internalization into germinated spores of Gigaspora margarita of two recombinant TAT fusion proteins consisting of either a fluorescent (GFP) or a luminescent (aequorin) reporter linked to the TAT peptide. Both TAT-fused proteins were found to enter AM fungal mycelia after a short incubation period (5-10 min). Ca2+ measurements in G. margarita mycelia pre-incubated with TAT-aequorin demonstrated the occurrence of changes in the intracellular free Ca2+ concentration in response to relevant stimuli, such as touch, cold, salinity, and strigolactones, symbiosis-related plant signals. These data indicate that the cell-penetrating properties of the TAT peptide can be used as an effective strategy for intracellularly delivering proteins of interest and shed new light on Ca2+ homeostasis and signalling in AM fungi.</AbstractText><CopyrightInformation>© 2014 The Authors. New Phytologist © 2014 New Phytologist Trust.</CopyrightInformation></Abstract><AuthorList CompleteYN="Y"><Author ValidYN="Y"><LastName>Moscatiello</LastName><ForeName>Roberto</ForeName><Initials>R</Initials><AffiliationInfo><Affiliation>Department of Biology, University of Padova, Via U. 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Phytol</MedlineTA><NlmUniqueID>9882884</NlmUniqueID><ISSNLinking>0028-646X</ISSNLinking></MedlineJournalInfo><ChemicalList><Chemical><RegistryNumber>0</RegistryNumber><NameOfSubstance UI="C507382">GR24 compound</NameOfSubstance></Chemical><Chemical><RegistryNumber>0</RegistryNumber><NameOfSubstance UI="D007783">Lactones</NameOfSubstance></Chemical><Chemical><RegistryNumber>0</RegistryNumber><NameOfSubstance UI="D010455">Peptides</NameOfSubstance></Chemical><Chemical><RegistryNumber>0</RegistryNumber><NameOfSubstance UI="D054322">tat Gene Products, Human Immunodeficiency Virus</NameOfSubstance></Chemical><Chemical><RegistryNumber>147336-22-9</RegistryNumber><NameOfSubstance UI="D049452">Green Fluorescent Proteins</NameOfSubstance></Chemical><Chemical><RegistryNumber>50934-79-7</RegistryNumber><NameOfSubstance UI="D000331">Aequorin</NameOfSubstance></Chemical><Chemical><RegistryNumber>SY7Q814VUP</RegistryNumber><NameOfSubstance UI="D002118">Calcium</NameOfSubstance></Chemical></ChemicalList><CitationSubset>IM</CitationSubset><MeshHeadingList><MeshHeading><DescriptorName UI="D000331" MajorTopicYN="N">Aequorin</DescriptorName><QualifierName UI="Q000378" MajorTopicYN="Y">metabolism</QualifierName></MeshHeading><MeshHeading><DescriptorName UI="D002118" MajorTopicYN="N">Calcium</DescriptorName><QualifierName UI="Q000378" MajorTopicYN="Y">metabolism</QualifierName></MeshHeading><MeshHeading><DescriptorName UI="D004705" MajorTopicYN="N">Endocytosis</DescriptorName><QualifierName UI="Q000187" MajorTopicYN="N">drug effects</QualifierName></MeshHeading><MeshHeading><DescriptorName UI="D004777" MajorTopicYN="N">Environment</DescriptorName></MeshHeading><MeshHeading><DescriptorName UI="D018014" MajorTopicYN="Y">Gene Transfer Techniques</DescriptorName></MeshHeading><MeshHeading><DescriptorName UI="D055137" MajorTopicYN="N">Glomeromycota</DescriptorName><QualifierName UI="Q000187" MajorTopicYN="N">drug 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UI="Q000494" MajorTopicYN="N">pharmacology</QualifierName></MeshHeading><MeshHeading><DescriptorName UI="D008163" MajorTopicYN="N">Luminescent Measurements</DescriptorName></MeshHeading><MeshHeading><DescriptorName UI="D038821" MajorTopicYN="N">Mycorrhizae</DescriptorName><QualifierName UI="Q000187" MajorTopicYN="N">drug effects</QualifierName><QualifierName UI="Q000502" MajorTopicYN="Y">physiology</QualifierName></MeshHeading><MeshHeading><DescriptorName UI="D010455" MajorTopicYN="N">Peptides</DescriptorName><QualifierName UI="Q000378" MajorTopicYN="N">metabolism</QualifierName></MeshHeading><MeshHeading><DescriptorName UI="D013559" MajorTopicYN="N">Symbiosis</DescriptorName><QualifierName UI="Q000187" MajorTopicYN="N">drug effects</QualifierName><QualifierName UI="Q000502" MajorTopicYN="Y">physiology</QualifierName></MeshHeading><MeshHeading><DescriptorName UI="D054322" MajorTopicYN="N">tat Gene Products, Human Immunodeficiency Virus</DescriptorName><QualifierName UI="Q000378" MajorTopicYN="Y">metabolism</QualifierName></MeshHeading></MeshHeadingList><KeywordList Owner="NOTNLM"><Keyword MajorTopicYN="N">Gigaspora margarita</Keyword><Keyword MajorTopicYN="N">TAT-GFP</Keyword><Keyword MajorTopicYN="N">TAT-aequorin</Keyword><Keyword MajorTopicYN="N">arbuscular mycorrhizal (AM) fungi</Keyword><Keyword MajorTopicYN="N">calcium</Keyword><Keyword MajorTopicYN="N">cell-penetrating peptides</Keyword><Keyword MajorTopicYN="N">strigolactones</Keyword></KeywordList></MedlineCitation><PubmedData><History><PubMedPubDate PubStatus="received"><Year>2013</Year><Month>12</Month><Day>20</Day></PubMedPubDate><PubMedPubDate PubStatus="accepted"><Year>2014</Year><Month>04</Month><Day>10</Day></PubMedPubDate><PubMedPubDate PubStatus="entrez"><Year>2014</Year><Month>5</Month><Day>22</Day><Hour>6</Hour><Minute>0</Minute></PubMedPubDate><PubMedPubDate PubStatus="pubmed"><Year>2014</Year><Month>5</Month><Day>23</Day><Hour>6</Hour><Minute>0</Minute></PubMedPubDate><PubMedPubDate 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