A GAF-domain-regulated adenylyl cyclase from Anabaena is a self-activating cAMP switch
The gene cyaB1 from the cyanobacterium Anabaena sp. PCC 7120 codes for a protein consisting of two N‐terminal GAF domains (GAF‐A and GAF‐B), a PAS domain and a class III adenylyl cyclase catalytic domain. The catalytic domain is active as a homodimer, as demonstrated by reconstitution from complemen...
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| Veröffentlicht in: | The EMBO journal 2002-07, Vol.21 (14), p.3672-3680 |
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| creator | Kanacher, Tobias Schultz, Anita Linder, Jürgen U. Schultz, Joachim E. |
| description | The gene
cyaB1
from the cyanobacterium
Anabaena
sp. PCC 7120 codes for a protein consisting of two N‐terminal GAF domains (GAF‐A and GAF‐B), a PAS domain and a class III adenylyl cyclase catalytic domain. The catalytic domain is active as a homodimer, as demonstrated by reconstitution from complementary inactive point mutants. The specific activity of the holoenyzme increased exponentially with time because the product cAMP activated dose dependently and nucleotide specifically (half‐maximally at 1 μM), identifying cAMP as a novel GAF domain ligand. Using point mutants of either the GAF‐A or GAF‐B domain revealed that cAMP activated via the GAF‐B domain. We replaced the cyanobacterial GAF domain ensemble in cyaB1 with the tandem GAF‐A/GAF‐B assemblage from the rat cGMP‐stimulated phosphodiesterase type 2, and converted cyaB1 to a cGMP‐stimulated adenylyl cyclase. This demonstrated the functional conservation of the GAF domain ensemble since the divergence of bacterial and eukaryotic lineages >2 billion years ago. In cyanobacteria, cyaB1 may act as a cAMP switch to stabilize committed developmental decisions. |
| doi_str_mv | 10.1093/emboj/cdf375 |
| format | Article |
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cyaB1
from the cyanobacterium
Anabaena
sp. PCC 7120 codes for a protein consisting of two N‐terminal GAF domains (GAF‐A and GAF‐B), a PAS domain and a class III adenylyl cyclase catalytic domain. The catalytic domain is active as a homodimer, as demonstrated by reconstitution from complementary inactive point mutants. The specific activity of the holoenyzme increased exponentially with time because the product cAMP activated dose dependently and nucleotide specifically (half‐maximally at 1 μM), identifying cAMP as a novel GAF domain ligand. Using point mutants of either the GAF‐A or GAF‐B domain revealed that cAMP activated via the GAF‐B domain. We replaced the cyanobacterial GAF domain ensemble in cyaB1 with the tandem GAF‐A/GAF‐B assemblage from the rat cGMP‐stimulated phosphodiesterase type 2, and converted cyaB1 to a cGMP‐stimulated adenylyl cyclase. This demonstrated the functional conservation of the GAF domain ensemble since the divergence of bacterial and eukaryotic lineages >2 billion years ago. In cyanobacteria, cyaB1 may act as a cAMP switch to stabilize committed developmental decisions.</description><identifier>ISSN: 0261-4189</identifier><identifier>ISSN: 1460-2075</identifier><identifier>EISSN: 1460-2075</identifier><identifier>DOI: 10.1093/emboj/cdf375</identifier><identifier>PMID: 12110580</identifier><identifier>CODEN: EMJODG</identifier><language>eng</language><publisher>Chichester, UK: John Wiley & Sons, Ltd</publisher><subject>Adenylate Cyclase Toxin ; adenylyl cyclases ; Adenylyl Cyclases - chemistry ; Adenylyl Cyclases - metabolism ; Amino Acid Sequence ; Anabaena ; Anabaena - enzymology ; Bacterial Proteins - metabolism ; Catalytic Domain ; Cyclic AMP - metabolism ; EMBO23 ; EMBO37 ; evolution ; GAF domain ; Molecular Sequence Data ; PAS domain ; Protein Precursors - metabolism ; Sequence Homology, Amino Acid</subject><ispartof>The EMBO journal, 2002-07, Vol.21 (14), p.3672-3680</ispartof><rights>European Molecular Biology Organization 2002</rights><rights>Copyright © 2002 European Molecular Biology Organization</rights><rights>Copyright Oxford University Press(England) Jul 15, 2002</rights><rights>Copyright © 2002 European Molecular Biology Organization 2002</rights><lds50>peer_reviewed</lds50><oa>free_for_read</oa><woscitedreferencessubscribed>false</woscitedreferencessubscribed><citedby>FETCH-LOGICAL-c6474-fe8a324e35a4a9031123e4ad8fa3f51481137e67d0994f62682054d671e7fccf3</citedby></display><links><openurl>$$Topenurl_article</openurl><openurlfulltext>$$Topenurlfull_article</openurlfulltext><thumbnail>$$Tsyndetics_thumb_exl</thumbnail><linktopdf>$$Uhttps://www.ncbi.nlm.nih.gov/pmc/articles/PMC126120/pdf/$$EPDF$$P50$$Gpubmedcentral$$H</linktopdf><linktohtml>$$Uhttps://www.ncbi.nlm.nih.gov/pmc/articles/PMC126120/$$EHTML$$P50$$Gpubmedcentral$$H</linktohtml><link.rule.ids>230,314,727,780,784,885,1416,1432,27923,27924,45573,45574,46408,46832,53790,53792</link.rule.ids><backlink>$$Uhttps://www.ncbi.nlm.nih.gov/pubmed/12110580$$D View this record in MEDLINE/PubMed$$Hfree_for_read</backlink></links><search><creatorcontrib>Kanacher, Tobias</creatorcontrib><creatorcontrib>Schultz, Anita</creatorcontrib><creatorcontrib>Linder, Jürgen U.</creatorcontrib><creatorcontrib>Schultz, Joachim E.</creatorcontrib><title>A GAF-domain-regulated adenylyl cyclase from Anabaena is a self-activating cAMP switch</title><title>The EMBO journal</title><addtitle>EMBO J</addtitle><addtitle>EMBO J</addtitle><description>The gene
cyaB1
from the cyanobacterium
Anabaena
sp. PCC 7120 codes for a protein consisting of two N‐terminal GAF domains (GAF‐A and GAF‐B), a PAS domain and a class III adenylyl cyclase catalytic domain. The catalytic domain is active as a homodimer, as demonstrated by reconstitution from complementary inactive point mutants. The specific activity of the holoenyzme increased exponentially with time because the product cAMP activated dose dependently and nucleotide specifically (half‐maximally at 1 μM), identifying cAMP as a novel GAF domain ligand. Using point mutants of either the GAF‐A or GAF‐B domain revealed that cAMP activated via the GAF‐B domain. We replaced the cyanobacterial GAF domain ensemble in cyaB1 with the tandem GAF‐A/GAF‐B assemblage from the rat cGMP‐stimulated phosphodiesterase type 2, and converted cyaB1 to a cGMP‐stimulated adenylyl cyclase. This demonstrated the functional conservation of the GAF domain ensemble since the divergence of bacterial and eukaryotic lineages >2 billion years ago. In cyanobacteria, cyaB1 may act as a cAMP switch to stabilize committed developmental decisions.</description><subject>Adenylate Cyclase Toxin</subject><subject>adenylyl cyclases</subject><subject>Adenylyl Cyclases - chemistry</subject><subject>Adenylyl Cyclases - metabolism</subject><subject>Amino Acid Sequence</subject><subject>Anabaena</subject><subject>Anabaena - enzymology</subject><subject>Bacterial Proteins - metabolism</subject><subject>Catalytic Domain</subject><subject>Cyclic AMP - metabolism</subject><subject>EMBO23</subject><subject>EMBO37</subject><subject>evolution</subject><subject>GAF domain</subject><subject>Molecular Sequence Data</subject><subject>PAS domain</subject><subject>Protein Precursors - metabolism</subject><subject>Sequence Homology, Amino Acid</subject><issn>0261-4189</issn><issn>1460-2075</issn><issn>1460-2075</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>2002</creationdate><recordtype>article</recordtype><sourceid>EIF</sourceid><sourceid>8G5</sourceid><sourceid>ABUWG</sourceid><sourceid>AFKRA</sourceid><sourceid>AZQEC</sourceid><sourceid>BENPR</sourceid><sourceid>CCPQU</sourceid><sourceid>DWQXO</sourceid><sourceid>GNUQQ</sourceid><sourceid>GUQSH</sourceid><sourceid>M2O</sourceid><recordid>eNqFkctv1DAQxiMEotvCjSso4tAToXZsx8mBw1J1F9CWh8SjN2vWGW-9TZxiJy373-OS1VIeAsmSLc3vmxl_X5I8ouQ5JRU7wnbZrY90bZgUd5IJ5QXJciLF3WRC8oJmnJbVXrIfwpoQIkpJ7yd7NKc0vskk-TxN59NZVnctWJd5XA0N9FinUKPbNJsm1RvdQMDU-K5Npw6WgA5SG1JIAzYmA93bK-itW6V6evo-Dde21-cPknsGmoAPt_dB8ml28vH4VbZ4N399PF1kuuCSZwZLYDlHJoBDRRilOUMOdWmAGUF5SSmTWMiaVBU3RV6UORG8LiRFabQ27CB5Mfa9HJYt1hpd76FRl9624DeqA6t-rTh7rlbdlaLRmpxE_eFW77uvA4ZetTZobBpw2A1ByWgekVX1X5CWnFHBRQSf_gauu8G7aIKilcgFJ7KM0LMR0r4LwaPZbUyJuklV_UhVjalG_MntX_6EtzFGQIzAtW1w889m6uT05RspKh5P1GWjLkSJW6G_tezfF3k88g76weNu0B_9bOjx264M_kIV8mb4l7dzJWYfzmbFGVUL9h0eFNfl</recordid><startdate>20020715</startdate><enddate>20020715</enddate><creator>Kanacher, Tobias</creator><creator>Schultz, Anita</creator><creator>Linder, Jürgen U.</creator><creator>Schultz, Joachim E.</creator><general>John Wiley & Sons, Ltd</general><general>Nature Publishing Group UK</general><general>Blackwell Publishing Ltd</general><general>Oxford University Press</general><scope>BSCLL</scope><scope>CGR</scope><scope>CUY</scope><scope>CVF</scope><scope>ECM</scope><scope>EIF</scope><scope>NPM</scope><scope>AAYXX</scope><scope>CITATION</scope><scope>3V.</scope><scope>7QG</scope><scope>7QL</scope><scope>7QP</scope><scope>7T5</scope><scope>7TK</scope><scope>7TM</scope><scope>7TO</scope><scope>7U9</scope><scope>7X7</scope><scope>7XB</scope><scope>88A</scope><scope>88E</scope><scope>8AO</scope><scope>8C1</scope><scope>8FD</scope><scope>8FE</scope><scope>8FH</scope><scope>8FI</scope><scope>8FJ</scope><scope>8FK</scope><scope>8G5</scope><scope>ABUWG</scope><scope>AEUYN</scope><scope>AFKRA</scope><scope>AZQEC</scope><scope>BBNVY</scope><scope>BENPR</scope><scope>BHPHI</scope><scope>BKSAR</scope><scope>C1K</scope><scope>CCPQU</scope><scope>DWQXO</scope><scope>FR3</scope><scope>FYUFA</scope><scope>GHDGH</scope><scope>GNUQQ</scope><scope>GUQSH</scope><scope>H94</scope><scope>HCIFZ</scope><scope>K9.</scope><scope>LK8</scope><scope>M0S</scope><scope>M1P</scope><scope>M2O</scope><scope>M7N</scope><scope>M7P</scope><scope>MBDVC</scope><scope>P64</scope><scope>PCBAR</scope><scope>PQEST</scope><scope>PQQKQ</scope><scope>PQUKI</scope><scope>Q9U</scope><scope>RC3</scope><scope>F1W</scope><scope>H95</scope><scope>H99</scope><scope>L.F</scope><scope>L.G</scope><scope>7X8</scope><scope>5PM</scope></search><sort><creationdate>20020715</creationdate><title>A GAF-domain-regulated adenylyl cyclase from Anabaena is a self-activating cAMP switch</title><author>Kanacher, Tobias ; Schultz, Anita ; Linder, Jürgen U. ; Schultz, Joachim E.</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-c6474-fe8a324e35a4a9031123e4ad8fa3f51481137e67d0994f62682054d671e7fccf3</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>2002</creationdate><topic>Adenylate Cyclase Toxin</topic><topic>adenylyl cyclases</topic><topic>Adenylyl Cyclases - chemistry</topic><topic>Adenylyl Cyclases - metabolism</topic><topic>Amino Acid Sequence</topic><topic>Anabaena</topic><topic>Anabaena - enzymology</topic><topic>Bacterial Proteins - metabolism</topic><topic>Catalytic Domain</topic><topic>Cyclic AMP - metabolism</topic><topic>EMBO23</topic><topic>EMBO37</topic><topic>evolution</topic><topic>GAF domain</topic><topic>Molecular Sequence Data</topic><topic>PAS domain</topic><topic>Protein Precursors - metabolism</topic><topic>Sequence Homology, Amino Acid</topic><toplevel>peer_reviewed</toplevel><toplevel>online_resources</toplevel><creatorcontrib>Kanacher, Tobias</creatorcontrib><creatorcontrib>Schultz, Anita</creatorcontrib><creatorcontrib>Linder, Jürgen U.</creatorcontrib><creatorcontrib>Schultz, Joachim E.</creatorcontrib><collection>Istex</collection><collection>Medline</collection><collection>MEDLINE</collection><collection>MEDLINE (Ovid)</collection><collection>MEDLINE</collection><collection>MEDLINE</collection><collection>PubMed</collection><collection>CrossRef</collection><collection>ProQuest Central (Corporate)</collection><collection>Animal Behavior Abstracts</collection><collection>Bacteriology Abstracts (Microbiology B)</collection><collection>Calcium & Calcified Tissue Abstracts</collection><collection>Immunology Abstracts</collection><collection>Neurosciences Abstracts</collection><collection>Nucleic Acids Abstracts</collection><collection>Oncogenes and Growth Factors Abstracts</collection><collection>Virology and AIDS Abstracts</collection><collection>Health & Medical Collection</collection><collection>ProQuest Central (purchase pre-March 2016)</collection><collection>Biology Database (Alumni Edition)</collection><collection>Medical Database (Alumni Edition)</collection><collection>ProQuest Pharma Collection</collection><collection>Public Health Database</collection><collection>Technology Research Database</collection><collection>ProQuest SciTech Collection</collection><collection>ProQuest Natural Science Collection</collection><collection>Hospital Premium Collection</collection><collection>Hospital Premium Collection (Alumni Edition)</collection><collection>ProQuest Central (Alumni) (purchase pre-March 2016)</collection><collection>Research Library (Alumni Edition)</collection><collection>ProQuest Central (Alumni Edition)</collection><collection>ProQuest One Sustainability</collection><collection>ProQuest Central UK/Ireland</collection><collection>ProQuest Central Essentials</collection><collection>Biological Science Collection</collection><collection>ProQuest Central</collection><collection>Natural Science Collection</collection><collection>Earth, Atmospheric & Aquatic Science Collection</collection><collection>Environmental Sciences and Pollution Management</collection><collection>ProQuest One Community College</collection><collection>ProQuest Central Korea</collection><collection>Engineering Research Database</collection><collection>Health Research Premium Collection</collection><collection>Health Research Premium Collection (Alumni)</collection><collection>ProQuest Central Student</collection><collection>Research Library Prep</collection><collection>AIDS and Cancer Research Abstracts</collection><collection>SciTech Premium Collection</collection><collection>ProQuest Health & Medical Complete (Alumni)</collection><collection>ProQuest Biological Science Collection</collection><collection>Health & Medical Collection (Alumni Edition)</collection><collection>Medical Database</collection><collection>Research Library</collection><collection>Algology Mycology and Protozoology Abstracts (Microbiology C)</collection><collection>Biological Science Database</collection><collection>Research Library (Corporate)</collection><collection>Biotechnology and BioEngineering Abstracts</collection><collection>Earth, Atmospheric & Aquatic Science Database</collection><collection>ProQuest One Academic Eastern Edition (DO NOT USE)</collection><collection>ProQuest One Academic</collection><collection>ProQuest One Academic UKI Edition</collection><collection>ProQuest Central Basic</collection><collection>Genetics Abstracts</collection><collection>ASFA: Aquatic Sciences and Fisheries Abstracts</collection><collection>Aquatic Science & Fisheries Abstracts (ASFA) 1: Biological Sciences & Living Resources</collection><collection>ASFA: Marine Biotechnology Abstracts</collection><collection>Aquatic Science & Fisheries Abstracts (ASFA) Marine Biotechnology Abstracts</collection><collection>Aquatic Science & Fisheries Abstracts (ASFA) Professional</collection><collection>MEDLINE - Academic</collection><collection>PubMed Central (Full Participant titles)</collection><jtitle>The EMBO journal</jtitle></facets><delivery><delcategory>Remote Search Resource</delcategory><fulltext>fulltext</fulltext></delivery><addata><au>Kanacher, Tobias</au><au>Schultz, Anita</au><au>Linder, Jürgen U.</au><au>Schultz, Joachim E.</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>A GAF-domain-regulated adenylyl cyclase from Anabaena is a self-activating cAMP switch</atitle><jtitle>The EMBO journal</jtitle><stitle>EMBO J</stitle><addtitle>EMBO J</addtitle><date>2002-07-15</date><risdate>2002</risdate><volume>21</volume><issue>14</issue><spage>3672</spage><epage>3680</epage><pages>3672-3680</pages><issn>0261-4189</issn><issn>1460-2075</issn><eissn>1460-2075</eissn><coden>EMJODG</coden><abstract>The gene
cyaB1
from the cyanobacterium
Anabaena
sp. PCC 7120 codes for a protein consisting of two N‐terminal GAF domains (GAF‐A and GAF‐B), a PAS domain and a class III adenylyl cyclase catalytic domain. The catalytic domain is active as a homodimer, as demonstrated by reconstitution from complementary inactive point mutants. The specific activity of the holoenyzme increased exponentially with time because the product cAMP activated dose dependently and nucleotide specifically (half‐maximally at 1 μM), identifying cAMP as a novel GAF domain ligand. Using point mutants of either the GAF‐A or GAF‐B domain revealed that cAMP activated via the GAF‐B domain. We replaced the cyanobacterial GAF domain ensemble in cyaB1 with the tandem GAF‐A/GAF‐B assemblage from the rat cGMP‐stimulated phosphodiesterase type 2, and converted cyaB1 to a cGMP‐stimulated adenylyl cyclase. This demonstrated the functional conservation of the GAF domain ensemble since the divergence of bacterial and eukaryotic lineages >2 billion years ago. In cyanobacteria, cyaB1 may act as a cAMP switch to stabilize committed developmental decisions.</abstract><cop>Chichester, UK</cop><pub>John Wiley & Sons, Ltd</pub><pmid>12110580</pmid><doi>10.1093/emboj/cdf375</doi><tpages>9</tpages><oa>free_for_read</oa></addata></record> |
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| source | MEDLINE; Wiley Free Content; EZB-FREE-00999 freely available EZB journals; Wiley Online Library All Journals; PubMed Central; Free Full-Text Journals in Chemistry |
| subjects | Adenylate Cyclase Toxin adenylyl cyclases Adenylyl Cyclases - chemistry Adenylyl Cyclases - metabolism Amino Acid Sequence Anabaena Anabaena - enzymology Bacterial Proteins - metabolism Catalytic Domain Cyclic AMP - metabolism EMBO23 EMBO37 evolution GAF domain Molecular Sequence Data PAS domain Protein Precursors - metabolism Sequence Homology, Amino Acid |
| title | A GAF-domain-regulated adenylyl cyclase from Anabaena is a self-activating cAMP switch |
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