Journal: The Journal of Biological Chemistry

Article Title: A Novel Interaction of the Catalytic Subunit of Protein Phosphatase 2A with the Adaptor Protein CIN85 Suppresses Phosphatase Activity and Facilitates Platelet Outside-in α IIb β 3 Integrin Signaling

doi: 10.1074/jbc.M115.704296

Figure Lengend Snippet: CIN85 is a novel PP2Ac interacting protein. A, lysate from HEK 293 cells transfected with either EV, HA-tagged PP2Ac and FLAG-tagged CIN85 or HA-tagged DUSP7 and FLAG-tagged CIN85 were immunoprecipitated (IP) with anti-FLAG antibody and immunoblotted (IB) with anti-HA and FLAG antibodies. Lysate panel shows the expression of PP2Ac (∼36 kDa), DUSP7 (∼44 kDa), and CIN85 (∼85 kDa). Blots are representative of 3 experiments. B, characterization of GST proteins by Coomassie Blue staining. C, CIN85-GST or GST was coupled to glutathione beads and used for a pulldown assays with purified PP2Ac and recombinant VWF A1A2A3 domain protein. PP2Ac and VWF A1A2A3 domain protein used in pulldown assays are depicted as input. Blots are representative of 3 experiments. D, increasing concentrations of CIN85-GST were added to immobilized PP2Ac; the CIN85-GST protein bound to PP2Ac was detected using ELISA. *, p < 0.05. n = 3. E, increasing concentrations of CIN85-GST and GST proteins were incubated with immobilized PP2Ac and the resultant phosphatase activity was assayed using a PP2A phosphatase activity kit. n = 3.

Article Snippet: Cells transfected with empty FLAG and HA vectors (EV), HA-tagged dual-specificity phosphatase 7 (DUSP7) (gift from Igor Astsaturov; Addgene plasmid #27976), and FLAG-tagged CIN85 served as controls.

Techniques: Transfection, Immunoprecipitation, Expressing, Staining, Purification, Recombinant, Enzyme-linked Immunosorbent Assay, Incubation, Activity Assay

Journal: The ISME Journal

Article Title: Is there a common water-activity limit for the three domains of life?

doi: 10.1038/ismej.2014.219

Figure Lengend Snippet: Growth curves for halophilic Bacteria and Archaea cultured in high-NaCl substrates and plotted in relation to water activity. ( a ) The bacterium Halorhodospira halochloris (strain and incubation temperature not specified) cultured in a defined medium supplemented with NaCl (calculated and replotted against water activity using data from Deole et al. , 2013 ). ( b ) The bacterium Halorhodospira halophila (strain DSM 244 T ; incubation temperature not specified) cultured in a defined medium supplemented with NaCl (calculated and replotted against water activity using data from Deole et al. , 2013 ). ( c ) The bacterium Halanaerobium lacusrosei (strain DSM 10165 T ) cultured in a complex medium supplemented with NaCl and incubated at 37 °C (calculated and replotted against water activity using data from Cayol et al. , 1995 ). ( d ) The Aacterium Actinopolyspora halophila (strain ATCC 27976 T ) cultured in a complex medium supplemented with NaCl, after a 14-day incubation at 37 °C (calculated and replotted against water activity using data from Yoshida et al. , 1991 ). ( e ) The Archaea ‘Haloarcula californiae' (strain DSM 8905; black line) and ‘Haloarcula sinaiiensis' (strain DSM 8928; orange line) cultured in a complex medium supplemented with NaCl and incubated at 37 °C (calculated and replotted against water activity using data from Javor et al. , 1982 ). ( f ) The archaeon Halorhabdus utahensis (strain DSM 12940 T ) in a defined medium supplemented with NaCl and incubated at 30 °C (calculated and re-plotted against water activity using data from Wainø et al. , 2000 ). For all media, water activity values were determined as described in the Materials and methods and at the same temperature as incubation was carried out for each set of media. Curves were extrapolated via regression analyses (dotted lines; for details see ) in order to determine the theoretical water activity minima for growth. Pink dashed lines indicate the previously accepted water activity limit for extremely halophilic Bacteria and Archaea (see ; ; Kminek et al. , 2010 ).

Article Snippet: Data obtained were compared with those for Haloquadratum walsbyi (DSM 16790), Pontibacillus (strain AS2), Salinicola (strain LC26), haloarchaeal strains GN-2 and GN-5, Halorhodospira halochloris , Halorhodospira halophila strain (DSM 244 T ), Halanaerobium lacusrosei (strain DSM 10165 T ), Actinopolyspora halophila (strain ATCC 27976 T ), ‘Haloarcula californiae' (strain DSM 8905), ‘Haloarcula sinaiiensis' (strain DSM 8928) and Halorhabdus utahensis (strain DSM 12940 T ); see ).

Techniques: Cell Culture, Activity Assay, Incubation

Journal: The ISME Journal

Article Title: Is there a common water-activity limit for the three domains of life?

doi: 10.1038/ismej.2014.219

Figure Lengend Snippet: Lower water-activity limits for cell division of the most xerophilic eukaryotic microbes (upper pale-blue panel) and Bacteria and Archaea thus far identified (lower pale-grey panel) on salt-supplemented substrates (mid-blue and dark-grey bars, respectively) and sugar- or polyol-supplemented substrates (dark-blue and mid-grey bars, respectively): (i) haloarchaeal strain GN-5 , (ii) haloarchaeal strain GN-2 , (iii) Halorhodospira halophila (strain DSM 244 T ; ), (iv) Halorhabdus utahensis (strain DSM 12940 T ; ), (v) Halobacterium strain 004.1 , (vi) Actinopolyspora halophila (strain ATCC 27976 T ; ), (vii) Halanaerobium lacusrosei (strain DSM 10165 T ; ), (viii) Halorhodospira halochloris (strain not specified; ), (ix) Bacteria within a mixed halophile community , (x) Natrinema pallidum (strain NCIMB 777 T ; and ), (xi) Halobacterium noricense (strain DSM 15987 T ; and ), (xii) Halcococcus salifodinae (strain DSM 13046; and ), (xiii) ‘Haloarcula californiae' (strain DSM 8905; ), (xiv) Haloquadratum walsbyi (strain DSM 16790; and ), (xv) ‘Haloarcula sinaiiensis' (strain DSM 8928; ), [xvi) Mycobacterium parascrofulaceum (strain LAIST_NPS017), (xvii) Mycobacterium smegmatis (strain ATCC 10143), (xviii) Tetragenococcus halophilus (strains T11 and T15), (xix) Saccharibacter floricola (strain DSM 15669 T ), (xx) Staphylococcus aureus (strains ATCC 6538P, NA and FM1), (xxi) Asaia bogorensis (strain JCM 10569 T ), (xxii) Gluconacetobacter diazotrophicus (strain DSM 5601 T ), (xxiii) Streptomyces albidoflavus (strain JCM 4198 T ), (xxiv) Staphylococcus epidermidis , (xxv) Halotalea alkalilenta (strain AW-7 T ), (xxvi) Streptomyces rectiviolaceus (strain JCM 9092 T ), (xxvii) Micromonospora grisea (strain JCM 3182), (xxviii) Sarcina sp. (strain 2b), (xxix) Lactococcus lactis (strain not specified), (xxx) Micromonospora sp. (strain JCM 3050), (see for (xvi) to (xxx); for (xxiii; xxvi; xxvii; xxx)), (xxxi) Basipetospora halophila (strain FRR 2787), (xxxii) Wallemia ichthyophaga (strain EXF-994), (xxxiii) Dunaliella salina (strain UTEX 200), (xxxiv) Polypaecilum pisce (strain FRR 2733), (xxxv) Dunaliella peircei (strain UTEX 2192), (xxxvi) Aspergillus penicilliodes (strain FRR 2612), (xxxvii) germination of Wallemia sebi (strain FRR 1473), (xxxviii) Eurotium halotolerans (strain EXF-4356), (xxxix) Halocafeteria seosinensis (strain EHF34), (xl) Dunaliella parva (strain UTEX 1983), (xli) Pleurostomum flabellatum (strain CCAP 1959/1), (xlii) Hortaea werneckii (strain EXF-225), (xliii) Euplaesiobystra hypersalinica (strain CCAP 1528/1), (xliv) Wallemia muriae (strain EXF-951), (xlv) Debaryomyces hansenii (strain DSM 70590); see for entries (xxxi) to (xlv), (xlvi) germination of Xeromyces bisporus (strain FRR 0025) on a watchglass in a humidity-controlled atmosphere , (xlvii) Zygosaccharomyces rouxii (strain not specified) on a high-sugar medium , (xlviii) germination of Aspergillus echinulatus (strain not specified) on a watchglass in a humidity-controlled atmosphere , (xlix) A. penicillioides (strain JH06THJ) , (l) X. bisporus (strain FRR 3443) , (li) Eurotium amstelodami (strains FRR 2792 and FRR 0475) on media supplemented with glycerol and other solutes , (lii) Eurotium chevalieri strain JH06THI , (liii) Xerochrysium xerophilium (formerly Chyrsosporium xerophilum Pitt et al. , 2013 ) (strain CBS 153.67 T ) on a medium supplemented with glucose and fructose ( Leong et al. , 2011 ), (liv) Eurotium repens (strain JH06JPD) on a medium supplemented with glycerol and other solutes , (lv) germination and growth of Eurotium halophilicum (strain FRR 2471) on a medium supplemented with glucose and fructose , (lvi) germination of Aspergillus penicillioides (strain not specified) in complex substrates ; (lvii) germination of Bettsia fastidia (formerly Chrysosporium fastidium , Pitt et al. , 2013 ) (strain FRR 77) on a watchglass in a humidity-controlled atmosphere , (lviii) germination of W. sebi (strain FRR 1473) on a medium supplemented with glucose and fructose , (lix) hyphal growth of B. fastidia (strain FRR 77) ( ; ), (lx) germination of Eurotium rubrum (strain FRR 0326) ( Gock et al. , 2003 ). For each bar, the shaded region extends to the lowest empirically determined water-activity value (see also and , , , ). Only lower water-activity limits for growth are indicated (unless spore germination is indicated); note that some of these species may be unable to grow close to a water activity of 1 (for examples, see ). The pink dashed line indicates the previously accepted water-activity limit for growth of the most halophilic Bacteria and Archaea (see ; ; Kminek et al. , 2010 ); the yellow dashed line indicates the original water-activity limit for hyphal growth of the most xerophilic fungi .

Article Snippet: Data obtained were compared with those for Haloquadratum walsbyi (DSM 16790), Pontibacillus (strain AS2), Salinicola (strain LC26), haloarchaeal strains GN-2 and GN-5, Halorhodospira halochloris , Halorhodospira halophila strain (DSM 244 T ), Halanaerobium lacusrosei (strain DSM 10165 T ), Actinopolyspora halophila (strain ATCC 27976 T ), ‘Haloarcula californiae' (strain DSM 8905), ‘Haloarcula sinaiiensis' (strain DSM 8928) and Halorhabdus utahensis (strain DSM 12940 T ); see ).

Techniques: Activity Assay

Journal: The ISME Journal

Article Title: Is there a common water-activity limit for the three domains of life?

doi: 10.1038/ismej.2014.219

Figure Lengend Snippet: Growth curves for halophilic Bacteria and Archaea cultured in high-NaCl substrates and plotted in relation to water activity. ( a ) The bacterium Halorhodospira halochloris (strain and incubation temperature not specified) cultured in a defined medium supplemented with NaCl (calculated and replotted against water activity using data from Deole et al. , 2013 ). ( b ) The bacterium Halorhodospira halophila (strain DSM 244 T ; incubation temperature not specified) cultured in a defined medium supplemented with NaCl (calculated and replotted against water activity using data from Deole et al. , 2013 ). ( c ) The bacterium Halanaerobium lacusrosei (strain DSM 10165 T ) cultured in a complex medium supplemented with NaCl and incubated at 37 °C (calculated and replotted against water activity using data from Cayol et al. , 1995 ). ( d ) The Aacterium Actinopolyspora halophila (strain ATCC 27976 T ) cultured in a complex medium supplemented with NaCl, after a 14-day incubation at 37 °C (calculated and replotted against water activity using data from Yoshida et al. , 1991 ). ( e ) The Archaea ‘Haloarcula californiae' (strain DSM 8905; black line) and ‘Haloarcula sinaiiensis' (strain DSM 8928; orange line) cultured in a complex medium supplemented with NaCl and incubated at 37 °C (calculated and replotted against water activity using data from Javor et al. , 1982 ). ( f ) The archaeon Halorhabdus utahensis (strain DSM 12940 T ) in a defined medium supplemented with NaCl and incubated at 30 °C (calculated and re-plotted against water activity using data from Wainø et al. , 2000 ). For all media, water activity values were determined as described in the Materials and methods and at the same temperature as incubation was carried out for each set of media. Curves were extrapolated via regression analyses (dotted lines; for details see ) in order to determine the theoretical water activity minima for growth. Pink dashed lines indicate the previously accepted water activity limit for extremely halophilic Bacteria and Archaea (see ; ; Kminek et al. , 2010 ).

Article Snippet: Data obtained were compared with those for Haloquadratum walsbyi (DSM 16790), Pontibacillus (strain AS2), Salinicola (strain LC26), haloarchaeal strains GN-2 and GN-5, Halorhodospira halochloris , Halorhodospira halophila strain (DSM 244 T ), Halanaerobium lacusrosei (strain DSM 10165 T ), Actinopolyspora halophila (strain ATCC 27976 T ), ‘Haloarcula californiae' (strain DSM 8905), ‘Haloarcula sinaiiensis' (strain DSM 8928) and Halorhabdus utahensis (strain DSM 12940 T ); see ).

Techniques: Cell Culture, Activity Assay, Incubation

Journal: The ISME Journal

Article Title: Is there a common water-activity limit for the three domains of life?

doi: 10.1038/ismej.2014.219

Figure Lengend Snippet: Lower water-activity limits for cell division of the most xerophilic eukaryotic microbes (upper pale-blue panel) and Bacteria and Archaea thus far identified (lower pale-grey panel) on salt-supplemented substrates (mid-blue and dark-grey bars, respectively) and sugar- or polyol-supplemented substrates (dark-blue and mid-grey bars, respectively): (i) haloarchaeal strain GN-5 , (ii) haloarchaeal strain GN-2 , (iii) Halorhodospira halophila (strain DSM 244 T ; ), (iv) Halorhabdus utahensis (strain DSM 12940 T ; ), (v) Halobacterium strain 004.1 , (vi) Actinopolyspora halophila (strain ATCC 27976 T ; ), (vii) Halanaerobium lacusrosei (strain DSM 10165 T ; ), (viii) Halorhodospira halochloris (strain not specified; ), (ix) Bacteria within a mixed halophile community , (x) Natrinema pallidum (strain NCIMB 777 T ; and ), (xi) Halobacterium noricense (strain DSM 15987 T ; and ), (xii) Halcococcus salifodinae (strain DSM 13046; and ), (xiii) ‘Haloarcula californiae' (strain DSM 8905; ), (xiv) Haloquadratum walsbyi (strain DSM 16790; and ), (xv) ‘Haloarcula sinaiiensis' (strain DSM 8928; ), [xvi) Mycobacterium parascrofulaceum (strain LAIST_NPS017), (xvii) Mycobacterium smegmatis (strain ATCC 10143), (xviii) Tetragenococcus halophilus (strains T11 and T15), (xix) Saccharibacter floricola (strain DSM 15669 T ), (xx) Staphylococcus aureus (strains ATCC 6538P, NA and FM1), (xxi) Asaia bogorensis (strain JCM 10569 T ), (xxii) Gluconacetobacter diazotrophicus (strain DSM 5601 T ), (xxiii) Streptomyces albidoflavus (strain JCM 4198 T ), (xxiv) Staphylococcus epidermidis , (xxv) Halotalea alkalilenta (strain AW-7 T ), (xxvi) Streptomyces rectiviolaceus (strain JCM 9092 T ), (xxvii) Micromonospora grisea (strain JCM 3182), (xxviii) Sarcina sp. (strain 2b), (xxix) Lactococcus lactis (strain not specified), (xxx) Micromonospora sp. (strain JCM 3050), (see for (xvi) to (xxx); for (xxiii; xxvi; xxvii; xxx)), (xxxi) Basipetospora halophila (strain FRR 2787), (xxxii) Wallemia ichthyophaga (strain EXF-994), (xxxiii) Dunaliella salina (strain UTEX 200), (xxxiv) Polypaecilum pisce (strain FRR 2733), (xxxv) Dunaliella peircei (strain UTEX 2192), (xxxvi) Aspergillus penicilliodes (strain FRR 2612), (xxxvii) germination of Wallemia sebi (strain FRR 1473), (xxxviii) Eurotium halotolerans (strain EXF-4356), (xxxix) Halocafeteria seosinensis (strain EHF34), (xl) Dunaliella parva (strain UTEX 1983), (xli) Pleurostomum flabellatum (strain CCAP 1959/1), (xlii) Hortaea werneckii (strain EXF-225), (xliii) Euplaesiobystra hypersalinica (strain CCAP 1528/1), (xliv) Wallemia muriae (strain EXF-951), (xlv) Debaryomyces hansenii (strain DSM 70590); see for entries (xxxi) to (xlv), (xlvi) germination of Xeromyces bisporus (strain FRR 0025) on a watchglass in a humidity-controlled atmosphere , (xlvii) Zygosaccharomyces rouxii (strain not specified) on a high-sugar medium , (xlviii) germination of Aspergillus echinulatus (strain not specified) on a watchglass in a humidity-controlled atmosphere , (xlix) A. penicillioides (strain JH06THJ) , (l) X. bisporus (strain FRR 3443) , (li) Eurotium amstelodami (strains FRR 2792 and FRR 0475) on media supplemented with glycerol and other solutes , (lii) Eurotium chevalieri strain JH06THI , (liii) Xerochrysium xerophilium (formerly Chyrsosporium xerophilum Pitt et al. , 2013 ) (strain CBS 153.67 T ) on a medium supplemented with glucose and fructose ( Leong et al. , 2011 ), (liv) Eurotium repens (strain JH06JPD) on a medium supplemented with glycerol and other solutes , (lv) germination and growth of Eurotium halophilicum (strain FRR 2471) on a medium supplemented with glucose and fructose , (lvi) germination of Aspergillus penicillioides (strain not specified) in complex substrates ; (lvii) germination of Bettsia fastidia (formerly Chrysosporium fastidium , Pitt et al. , 2013 ) (strain FRR 77) on a watchglass in a humidity-controlled atmosphere , (lviii) germination of W. sebi (strain FRR 1473) on a medium supplemented with glucose and fructose , (lix) hyphal growth of B. fastidia (strain FRR 77) ( ; ), (lx) germination of Eurotium rubrum (strain FRR 0326) ( Gock et al. , 2003 ). For each bar, the shaded region extends to the lowest empirically determined water-activity value (see also and , , , ). Only lower water-activity limits for growth are indicated (unless spore germination is indicated); note that some of these species may be unable to grow close to a water activity of 1 (for examples, see ). The pink dashed line indicates the previously accepted water-activity limit for growth of the most halophilic Bacteria and Archaea (see ; ; Kminek et al. , 2010 ); the yellow dashed line indicates the original water-activity limit for hyphal growth of the most xerophilic fungi .

Article Snippet: Data obtained were compared with those for Haloquadratum walsbyi (DSM 16790), Pontibacillus (strain AS2), Salinicola (strain LC26), haloarchaeal strains GN-2 and GN-5, Halorhodospira halochloris , Halorhodospira halophila strain (DSM 244 T ), Halanaerobium lacusrosei (strain DSM 10165 T ), Actinopolyspora halophila (strain ATCC 27976 T ), ‘Haloarcula californiae' (strain DSM 8905), ‘Haloarcula sinaiiensis' (strain DSM 8928) and Halorhabdus utahensis (strain DSM 12940 T ); see ).

Techniques: Activity Assay