Tyrosine Phosphorylation as a Widespread Regulatory Mechanism in Prokaryotes

Getz, Landon J.; Runte, Cameron S.; Rainey, Jan K.; Thomas, Nikhil A.

doi:10.1128/jb.00205-19

Cited by 24 publications

(12 citation statements)

References 86 publications

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“…Additionally, we found that Tyr153 but not Tyr152 is the major phosphorylation site on CesT being phosphorylated by NleH2. This is contradictory to the previous finding involving molecular dynamic simulations for CsrA with phosphorylated CesT (Tyr152), which indicated that competitive binding: PO 3 ‐mediated tripartite H‐bonding between CseT and CsrA at specific residues (Ser41, Val42, and His43) could compete with CsrA binding to the “GGA” motif on target mRNA 28 . However, it was shown that Y153F mutation leads to increased Non‐LEE encoded effector A (NleA) translation and secretion 27 .…”

Section: Discussioncontrasting

confidence: 98%

“…This is contradictory to the previous finding involving molecular dynamic simulations for CsrA with phosphorylated CesT (Tyr152), which indicated that competitive binding: PO 3 -mediated tripartite H-bonding between CseT and CsrA at specific residues (Ser41, Val42, and His43) could compete with CsrA binding to the "GGA" motif on target mRNA. 28 However, it was shown that Y153F mutation leads to increased Non-LEE encoded effector A (NleA) translation and secretion. 27 This suggested that differential phosphorylation of CesT at Tyr152 and Tyr153 by NleH2 might be involved in regulating the translation and secretion of another effector protein NleA.…”

Section: Discussionmentioning

confidence: 99%

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Binding specificity of type three secretion system effector NleH2 to multi‐cargo chaperone CesT and their phosphorylation

et al. 2021

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Gram-negative pathogens like Enteropathogenic Escherichia coli (EPEC) utilize the type three secretion system (T3SS) to translocate various effector proteins that are needed to "hijack" the host system for pathogenic survival. Specialized T3SS chaperones inside bacterial cells stabilize these effector proteins and facilitate their translocation. CesT is a unique multi-cargo chaperone that interacts with and translocates ~10 different effector proteins. Here, we report the specific interaction between CesT and its key effector, NleH2, and explore the potential role of NleH2 as a kinase for CesT phosphorylation. First, we identified the chaperone-binding domain (CBD; 19-97aa) of NleH2, and mapped the specific interaction sites for both CesT and NleH2. The N-and Cterminal residues of the CBD interact with the dimeric interface of CesT. Further, we compared the CesT binding to NleH2, to that of another key effector Tir and with the global carbon regulator CsrA. Notably, the effectors have the binding regions at the β-sheet core and dimer interface of CesT, whereas the CsrA regulator interacts predominantly through the C-terminal region, which is found ~17 Å away from the effectors-binding sites. Next, we showed that NleH2 remains an active kinase even as a complex with CesT and is responsible for its autophosphorylation as well as phosphorylation of CesT at Tyr153. Collectively, our findings enhance the understanding of the role of multi-cargo chaperone CesT in orchestrating effector translocation through T3SS.

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Section: Discussioncontrasting

confidence: 98%

Section: Discussionmentioning

confidence: 99%

Binding specificity of type three secretion system effector NleH2 to multi‐cargo chaperone CesT and their phosphorylation

et al. 2021

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“…Readers are proteins without enzymatic activity that bind to specific protein domains once they carry a PTM, and mediate the effect of the modification in different ways, such as establishing or preventing protein interactions, or inducing a translocation of subcellular localisation (Figure 1B). The concept of writers, readers and erasers for posttranslational modifications, as described in [33], is known for a range of different modifications, such as the histone code in the field of epigenetics [34,35], or phosphotyrosine binding domains [36,37]. Also in glycobiology, a large number of glycosyltransferases (writers) and glycoside hydrolases (erasers) are complemented by proteins specifically recognizing different types of glycosylation (readers) [38,39].…”

Section: Accepted Articlementioning

confidence: 99%

Balancing O‐GlcNAc and O‐fucose in plants

Mutanwad

Lucyshyn

2021

The FEBS Journal

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O-linked modification of nuclear and cytosolic proteins with monosaccharides is essential in all eukaryotes.While many aspects of this post-translational modification are highly conserved, there are striking differences between plants and the animal kingdom. In animals, dynamic cycling of O-GlcNAc is established by two essential single copy enzymes, the O-GlcNAc transferase OGT and O-GlcNAc hydrolase OGA. In contrast, plants balance O-GlcNAc with O-fucose modifications, catalysed by the OGT SECRET AGENT (SEC) and the Protein O-Fucosyltransferase (POFUT) SPINDLY (SPY). However, specific glycoside hydrolases for either of the two modifications have not yet been identified. Nucleocytoplasmic O-glycosylation is still not very well understood in plants, even though a high number of proteins were found to be affected. One important open question is how specificity is established in a system where only two enzymes modify hundreds of proteins. Here, we discuss the possibility that O-GlcNAc and O-fucose binding proteins could introduce an additional flexible layer of regulation in Oglycosylation-mediated signaling pathways, with the potential of integrating internal or external signals. O-glycosylation of nucleocytoplasmic proteins -common features in different organisms Protein glycosylation is a very common post-translational modification (PTM), that comes in a lot of different shapes and sizes. The formation of classical N-and O-glycans usually results in branched structures that vary in size. Flexible further modifications such as addition, trimming or chemical modifications of the carbohydrates results in even higher diversity. Most of these glycosylated proteins are found in the secretory pathway, destined for extracellular or membrane-associated localisation, where glycans either assist protein folding or form important structural components of cell walls [1, 2]. Oglycosylation of nuclear and cytosolic proteins on the other hand falls in a completely different category: a single monosaccharide is O-linked to serine or threonine residues of the target proteins, without further attachment of additional sugar residues. The most prominent and best described example for this is O-GlcNAcylation [3-5], which was found in most eukaryotic organisms and some prokaryotes [6]. Classical O-GlcNAc modification is catalysed by the O-GlcNAc transferase (OGT), a highly conserved enzyme that uses UDP-GlcNAc as a donor substrate. In animals, the modification can be cleaved again by a specific O-GlcNAc hydrolase (OGA), which is -like OGT -a single copy enzyme. Both OGT and OGA are essential for animal development, as shown in several model organisms including Drosophila melanogaster [7, 8] and mice [9, 10]. A functional nucleocytoplasmic O-glycosyltransferase is also essential in plants, but a plant version of OGA has not yet been identified [6, 11].O-GlcNAc modifications were found in a wide range of different protein classes involved in many basic cellular pathways and signaling networks. Over numerous studies, more than 5000 human ...

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“…The phosphorylated tyrosine residue in EI, Tyr122, is a conserved residue between EI sugar and EI Ntr (33), suggestive of its importance. Notably, extensive tyrosine phosphorylation occurs in proteins linked to sugar metabolism and the TCA cycle in E. coli and in the archaeon Sulfolobus solfataricus (55), suggesting that the impact of tyrosine phosphorylation on proteins that function in carbohydrate metabolism is conserved through evolution. The finding that tyrosine phosphorylation regulates the activity of the catabolite regulator Cra (22), together with our findings that it regulates localization of EI and of the factor that controls EI activity, TmaR, supports this hypothesis.…”

Section: Coli Encounters Does Tmar Absence Confer a Disadvantage?mentioning

confidence: 99%

Tyrosine phosphorylation-dependent localization of TmaR, a novelE. colipolar protein that controls activity of the major sugar regulator by polar sequestration

Szoke

Albocher

Govindarajan

et al. 2020

Preprint

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The poles of E. coli cells are emerging as hubs for major sensory systems, but the polar determinants that allocate their components to the pole are largely unknown. Here, we describe the discovery of a novel protein, TmaR, which localizes to the E. coli cell pole when phosphorylated on a tyrosine residue. TmaR is shown here to control the subcellular localization of the general PTS protein Enzyme I (EI) by preventing it from exerting its activity by binding and polar sequestration, thus regulating sugar uptake and metabolism. Depletion or overexpression of TmaR results in EI release from the pole or enhanced recruitment to the pole, which leads to increasing or decreasing the rate of sugar consumption, respectively. Notably phosphorylation of TmaR is required to release EI and enable its activity. Like TmaR, the ability of EI to be recruited to the pole depends on phosphorylation of one of its tyrosines. In addition to hyperactivity in sugar consumption, the absence of TmaR also leads to detrimental effects on the ability of cells to survive in mild acidic conditions. Our results argue that this survival defect, which is sugar- and EI-dependent, reflects the difficulty of cells lacking TmaR to enter stationary phase. Our study identifies TmaR as the first E. coli protein reported to localize in a tyrosine-dependent manner and to control the activity of other proteins by their polar sequestration and release.SIGNIFICANCEIn recent years, we have learnt that bacterial cells have intricate spatial organization despite the lack of membrane-bounded organelles. The endcaps of rod-shaped bacteria, termed poles, are emerging as hubs for sensing and responding, but the underlying mechanisms for positioning macromolecules there are largely unknown. We discovered a novel protein, TmaR, whose polar localization depends on a phospho-tyrosine modification. We show that TmaR controls the activity of EI, the major regulator of sugar metabolism in most bacteria, by polar sequestration and release. Notably, TmaR is essential for survival in conditions that E. coli often encounters in nature. Hence, TmaR is a key regulator that connects tyrosine phosphorylation, spatial regulation, sugar metabolism and survival in bacteria and the first protein reported to recruit proteins to the E. coli cell poles.

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Tyrosine Phosphorylation as a Widespread Regulatory Mechanism in Prokaryotes

Cited by 24 publications

References 86 publications

Binding specificity of type three secretion system effector NleH2 to multi‐cargo chaperone CesT and their phosphorylation

Binding specificity of type three secretion system effector NleH2 to multi‐cargo chaperone CesT and their phosphorylation

Balancing O‐GlcNAc and O‐fucose in plants

Tyrosine phosphorylation-dependent localization of TmaR, a novelE. colipolar protein that controls activity of the major sugar regulator by polar sequestration

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