N-Glycosylation Is Important for Halobacterium salinarum Archaellin Expression, Archaellum Assembly and Cell Motility
Halobacterium salinarum are halophilic archaea that display directional swimming in response to various environmental signals, including light, chemicals and oxygen. In Hbt. salinarum, the building blocks (archaellins) of the archaeal swimming apparatus (the archaellum) are N-glycosylated. However,...
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| Format: | Article |
| Language: | English |
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Frontiers Media S.A.
2019-06-01
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| Series: | Frontiers in Microbiology |
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| Online Access: | https://www.frontiersin.org/article/10.3389/fmicb.2019.01367/full |
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| author | Marianna Zaretsky Cynthia L. Darnell Amy K. Schmid Amy K. Schmid Jerry Eichler |
| author_facet | Marianna Zaretsky Cynthia L. Darnell Amy K. Schmid Amy K. Schmid Jerry Eichler |
| author_sort | Marianna Zaretsky |
| collection | DOAJ |
| description | Halobacterium salinarum are halophilic archaea that display directional swimming in response to various environmental signals, including light, chemicals and oxygen. In Hbt. salinarum, the building blocks (archaellins) of the archaeal swimming apparatus (the archaellum) are N-glycosylated. However, the physiological importance of archaellin N-glycosylation remains unclear. Here, a tetrasaccharide comprising a hexose and three hexuronic acids decorating the five archaellins was characterized by mass spectrometry. Such analysis failed to detect sulfation of the hexuronic acids, in contrast to earlier reports. To better understand the physiological significance of Hbt. salinarum archaellin N-glycosylation, a strain deleted of aglB, encoding the archaeal oligosaccharyltransferase, was generated. In this ΔaglB strain, archaella were not detected and only low levels of archaellins were released into the medium, in contrast to what occurs with the parent strain. Mass spectrometry analysis of the archaellins in ΔaglB cultures did not detect N-glycosylation. ΔaglB cells also showed a slight growth defect and were impaired for motility. Quantitative real-time PCR analysis revealed dramatically reduced transcript levels of archaellin-encoding genes in the mutant strain, suggesting that N-glycosylation is important for archaellin transcription, with downstream effects on archaellum assembly and function. Control of AglB-dependent post-translational modification of archaellins could thus reflect a previously unrecognized route for regulating Hbt. salinarum motility. |
| first_indexed | 2024-12-22T03:54:48Z |
| format | Article |
| id | doaj.art-d342a1d1201549a588c899bf2ec71c1a |
| institution | Directory Open Access Journal |
| issn | 1664-302X |
| language | English |
| last_indexed | 2024-12-22T03:54:48Z |
| publishDate | 2019-06-01 |
| publisher | Frontiers Media S.A. |
| record_format | Article |
| series | Frontiers in Microbiology |
| spelling | doaj.art-d342a1d1201549a588c899bf2ec71c1a2022-12-21T18:39:54ZengFrontiers Media S.A.Frontiers in Microbiology1664-302X2019-06-011010.3389/fmicb.2019.01367468445N-Glycosylation Is Important for Halobacterium salinarum Archaellin Expression, Archaellum Assembly and Cell MotilityMarianna Zaretsky0Cynthia L. Darnell1Amy K. Schmid2Amy K. Schmid3Jerry Eichler4Department of Life Sciences, Ben Gurion University of the Negev, Beersheba, IsraelDepartment of Biology, Duke University, Durham, NC, United StatesDepartment of Biology, Duke University, Durham, NC, United StatesCenter for Genomics and Computational Biology, Duke University, Durham, NC, United StatesDepartment of Life Sciences, Ben Gurion University of the Negev, Beersheba, IsraelHalobacterium salinarum are halophilic archaea that display directional swimming in response to various environmental signals, including light, chemicals and oxygen. In Hbt. salinarum, the building blocks (archaellins) of the archaeal swimming apparatus (the archaellum) are N-glycosylated. However, the physiological importance of archaellin N-glycosylation remains unclear. Here, a tetrasaccharide comprising a hexose and three hexuronic acids decorating the five archaellins was characterized by mass spectrometry. Such analysis failed to detect sulfation of the hexuronic acids, in contrast to earlier reports. To better understand the physiological significance of Hbt. salinarum archaellin N-glycosylation, a strain deleted of aglB, encoding the archaeal oligosaccharyltransferase, was generated. In this ΔaglB strain, archaella were not detected and only low levels of archaellins were released into the medium, in contrast to what occurs with the parent strain. Mass spectrometry analysis of the archaellins in ΔaglB cultures did not detect N-glycosylation. ΔaglB cells also showed a slight growth defect and were impaired for motility. Quantitative real-time PCR analysis revealed dramatically reduced transcript levels of archaellin-encoding genes in the mutant strain, suggesting that N-glycosylation is important for archaellin transcription, with downstream effects on archaellum assembly and function. Control of AglB-dependent post-translational modification of archaellins could thus reflect a previously unrecognized route for regulating Hbt. salinarum motility.https://www.frontiersin.org/article/10.3389/fmicb.2019.01367/fullarchaeaarchaellinarchaellumHalobacterium salinarummotilityN-glycosylation |
| spellingShingle | Marianna Zaretsky Cynthia L. Darnell Amy K. Schmid Amy K. Schmid Jerry Eichler N-Glycosylation Is Important for Halobacterium salinarum Archaellin Expression, Archaellum Assembly and Cell Motility Frontiers in Microbiology archaea archaellin archaellum Halobacterium salinarum motility N-glycosylation |
| title | N-Glycosylation Is Important for Halobacterium salinarum Archaellin Expression, Archaellum Assembly and Cell Motility |
| title_full | N-Glycosylation Is Important for Halobacterium salinarum Archaellin Expression, Archaellum Assembly and Cell Motility |
| title_fullStr | N-Glycosylation Is Important for Halobacterium salinarum Archaellin Expression, Archaellum Assembly and Cell Motility |
| title_full_unstemmed | N-Glycosylation Is Important for Halobacterium salinarum Archaellin Expression, Archaellum Assembly and Cell Motility |
| title_short | N-Glycosylation Is Important for Halobacterium salinarum Archaellin Expression, Archaellum Assembly and Cell Motility |
| title_sort | n glycosylation is important for halobacterium salinarum archaellin expression archaellum assembly and cell motility |
| topic | archaea archaellin archaellum Halobacterium salinarum motility N-glycosylation |
| url | https://www.frontiersin.org/article/10.3389/fmicb.2019.01367/full |
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