Characterisation of Variants of Cyclic di-GMP Turnover Proteins Associated with Semi-Constitutive rdar Morphotype Expression in Commensal and Uropathogenic <i>Escherichia coli</i> Strains
Expression of rdar (red, dry, and rough) colony morphology-based biofilm formation in <i>Escherichia coli</i> is highly variable. To investigate the molecular mechanisms of semi-constitutive rdar morphotype formation, we compared their cyclic di-GMP turnover protein content and variabili...
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MDPI AG
2023-08-01
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| Series: | Microorganisms |
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| Online Access: | https://www.mdpi.com/2076-2607/11/8/2048 |
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| author | Annika Cimdins-Ahne Ali-Oddin Naemi Fengyang Li Roger Simm Ute Römling |
| author_facet | Annika Cimdins-Ahne Ali-Oddin Naemi Fengyang Li Roger Simm Ute Römling |
| author_sort | Annika Cimdins-Ahne |
| collection | DOAJ |
| description | Expression of rdar (red, dry, and rough) colony morphology-based biofilm formation in <i>Escherichia coli</i> is highly variable. To investigate the molecular mechanisms of semi-constitutive rdar morphotype formation, we compared their cyclic di-GMP turnover protein content and variability to the highly regulated, temperature-dependent morphotype of the historical and modern ST10 isolates <i>E. coli</i> MG1655 and Fec10, respectively. Subsequently, we assessed the effects of cyclic di-GMP turnover protein variants of the EAL phosphodiesterases YcgG and YjcC and the horizontally transferred diguanylate cyclase DgcX on biofilm formation and motility. The two YcgG variants with truncations of the N-terminal CSS signaling domain were oppositely effective in targeting downregulation of rdar biofilm formation compared to the full-length reference protein. Expression of the C-terminal truncated variants YjcC<sub>Fec67</sub> and YjcC<sub>Tob1</sub> showed highly diminished apparent phosphodiesterase activity compared to the reference YjcC<sub>MG1655</sub>. For YjcC<sub>Fec101</sub>, substitution of the C-terminus led to an apparently inactive enzyme. Overexpression of the diguanylate cyclase DgcX contributed to upregulation of cellulose biosynthesis but not to elevated expression of the major biofilm regulator <i>csgD</i> in the “classical” rdar-expressing commensal strain <i>E. coli</i> Fec10. Thus, the c-di-GMP regulating network is highly complex with protein variants displaying substantially different apparent enzymatic activities. |
| first_indexed | 2024-03-10T23:43:11Z |
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| language | English |
| last_indexed | 2024-03-10T23:43:11Z |
| publishDate | 2023-08-01 |
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| record_format | Article |
| series | Microorganisms |
| spelling | doaj.art-f6c573a8ee844c82a9540acbb21e1f9a2023-11-19T02:18:19ZengMDPI AGMicroorganisms2076-26072023-08-01118204810.3390/microorganisms11082048Characterisation of Variants of Cyclic di-GMP Turnover Proteins Associated with Semi-Constitutive rdar Morphotype Expression in Commensal and Uropathogenic <i>Escherichia coli</i> StrainsAnnika Cimdins-Ahne0Ali-Oddin Naemi1Fengyang Li2Roger Simm3Ute Römling4Department of Microbiology, Tumor and Cell Biology, Karolinska Institutet, 171 77 Stockholm, SwedenInstitute of Oral Biology, University of Oslo, 0313 Oslo, NorwayDepartment of Microbiology, Tumor and Cell Biology, Karolinska Institutet, 171 77 Stockholm, SwedenInstitute of Oral Biology, University of Oslo, 0313 Oslo, NorwayDepartment of Microbiology, Tumor and Cell Biology, Karolinska Institutet, 171 77 Stockholm, SwedenExpression of rdar (red, dry, and rough) colony morphology-based biofilm formation in <i>Escherichia coli</i> is highly variable. To investigate the molecular mechanisms of semi-constitutive rdar morphotype formation, we compared their cyclic di-GMP turnover protein content and variability to the highly regulated, temperature-dependent morphotype of the historical and modern ST10 isolates <i>E. coli</i> MG1655 and Fec10, respectively. Subsequently, we assessed the effects of cyclic di-GMP turnover protein variants of the EAL phosphodiesterases YcgG and YjcC and the horizontally transferred diguanylate cyclase DgcX on biofilm formation and motility. The two YcgG variants with truncations of the N-terminal CSS signaling domain were oppositely effective in targeting downregulation of rdar biofilm formation compared to the full-length reference protein. Expression of the C-terminal truncated variants YjcC<sub>Fec67</sub> and YjcC<sub>Tob1</sub> showed highly diminished apparent phosphodiesterase activity compared to the reference YjcC<sub>MG1655</sub>. For YjcC<sub>Fec101</sub>, substitution of the C-terminus led to an apparently inactive enzyme. Overexpression of the diguanylate cyclase DgcX contributed to upregulation of cellulose biosynthesis but not to elevated expression of the major biofilm regulator <i>csgD</i> in the “classical” rdar-expressing commensal strain <i>E. coli</i> Fec10. Thus, the c-di-GMP regulating network is highly complex with protein variants displaying substantially different apparent enzymatic activities.https://www.mdpi.com/2076-2607/11/8/2048rdar morphotypec-di-GMPbiofilmprotein variantsGGDEFEAL |
| spellingShingle | Annika Cimdins-Ahne Ali-Oddin Naemi Fengyang Li Roger Simm Ute Römling Characterisation of Variants of Cyclic di-GMP Turnover Proteins Associated with Semi-Constitutive rdar Morphotype Expression in Commensal and Uropathogenic <i>Escherichia coli</i> Strains Microorganisms rdar morphotype c-di-GMP biofilm protein variants GGDEF EAL |
| title | Characterisation of Variants of Cyclic di-GMP Turnover Proteins Associated with Semi-Constitutive rdar Morphotype Expression in Commensal and Uropathogenic <i>Escherichia coli</i> Strains |
| title_full | Characterisation of Variants of Cyclic di-GMP Turnover Proteins Associated with Semi-Constitutive rdar Morphotype Expression in Commensal and Uropathogenic <i>Escherichia coli</i> Strains |
| title_fullStr | Characterisation of Variants of Cyclic di-GMP Turnover Proteins Associated with Semi-Constitutive rdar Morphotype Expression in Commensal and Uropathogenic <i>Escherichia coli</i> Strains |
| title_full_unstemmed | Characterisation of Variants of Cyclic di-GMP Turnover Proteins Associated with Semi-Constitutive rdar Morphotype Expression in Commensal and Uropathogenic <i>Escherichia coli</i> Strains |
| title_short | Characterisation of Variants of Cyclic di-GMP Turnover Proteins Associated with Semi-Constitutive rdar Morphotype Expression in Commensal and Uropathogenic <i>Escherichia coli</i> Strains |
| title_sort | characterisation of variants of cyclic di gmp turnover proteins associated with semi constitutive rdar morphotype expression in commensal and uropathogenic i escherichia coli i strains |
| topic | rdar morphotype c-di-GMP biofilm protein variants GGDEF EAL |
| url | https://www.mdpi.com/2076-2607/11/8/2048 |
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