Nucleotide-dependent farnesyl switch orchestrates polymerization and membrane binding of human guanylate-binding protein 1

Shydlovskyi, Sergii; Zienert, Anke Y.; İnce, Semra; Dovengerds, Christine; Hohendahl, Annika; Dargazanli, Julia M.; Blum, Ailisa; Günther, Saskia Diana; Kladt, Nikolay; Stürzl, Michael; Schauß, Astrid; Kutsch, Miriam; Roux, Aurélien; Praefcke, Gerrit J. K.; Herrmann, Christian

doi:10.1073/pnas.1620959114

Cited by 61 publications

(144 citation statements)

References 56 publications

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“…This is not surprising when considering notable differences that the isoforms display at biochemical levels: for instance, hGBP-1 in clear contrast to hGBP-5 provides not only GMP-binding ability but also yields GMP from GTP hydrolysis [26,29]. In addition to that, we recently discovered that farnesylation also enables hGBP-1 to form polymers giving similar ring-like structures as observed for BDLP [63,64]. Prenylationi.e., farnesylation (hGBP-1) and geranylgeranylation (hGBP-5)is absolutely required for the proteins to associate with membranes [31,32].…”

Section: Discussionmentioning

confidence: 78%

The human guanylate‐binding proteins hGBP‐1 and hGBP‐5 cycle between monomers and dimers only

et al. 2017

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Belonging to the dynamin superfamily of large GTPases, human guanylate-binding proteins (hGBPs) comprise a family of seven isoforms (hGBP-1 to hGBP-7) that are strongly upregulated in response to interferon-γ and other cytokines. Accordingly, several hGBPs are found to exhibit various cellular functions encompassing inhibitory effects on cell proliferation, tumor suppression as well as antiviral and antibacterial activity; however, their mechanism of action is only poorly understood. Often, cellular functions of dynamin-related proteins are closely linked to their ability to form nucleotide-dependent oligomers, a feature that also applies to hGBP-1 and hGBP-5. hGBPs are described as monomers, dimers, tetramers, and higher oligomeric species, the function of which is not clearly established. Therefore, this work focused on the oligomerization capability of hGBP-1 and hGBP-5, which are reported to assemble to homodimers and homotetramers. Employing independent methods such as size-exclusion chromatography, which relies on the hydrodynamic radius, and multiangle light scattering, which relies on the mass of the protein, revealed that previous interpretations regarding the size of the proteins and their complexes have to be revised. Additional studies using inter- and intramolecular Förster resonance energy transfer demonstrated that nucleotide-triggered intramolecular structural changes lead to a more extended shape of hGBP-1 being responsible for the appearance of larger oligomeric species. Thus, previously reported tetrameric and dimeric species of hGBP-1 and hGBP-5 were unmasked as dimers and monomers, respectively, with their shapes depending on both the bound nucleotide and the ionic strength of the solution.

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Section: Discussionmentioning

confidence: 78%

The human guanylate‐binding proteins hGBP‐1 and hGBP‐5 cycle between monomers and dimers only

et al. 2017

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“…Interestingly, GBP membrane localization is usually transient and the three isoprenylated isoforms are able to further recruit nonprenylated GBPs to their respective localization. Moreover, cytoplasmic hGBP1 in HeLa cells has been shown to partially colocalize with proteins from the endolysosomal system, such as Rab5, EEA‐1, Rab7, and Lamp‐1 …”

Section: Introductionmentioning

confidence: 99%

Sensing of invading pathogens by GBPs: At the crossroads between cell-autonomous and innate immunity

Santos

Brož

2018

Journal of Leukocyte Biology

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Guanylate-binding proteins (GBPs) are conserved family of IFN-inducible GTPases that play an important role in the host immunity against bacterial, viral, and protozoan pathogens. GBPs protect the host by associating with intracellular microbes, their vacuolar niche or, in the case of viruses, with their replication complex. This association results in a restriction of the respective pathogen, yet the exact molecular mechanisms of the antimicrobial functions of GBPs are still unclear. Recent work has linked the GBPs with the activation of inflammasomes, multi-protein complexes that assemble upon recognition of pathogen- or host-derived signals and that drive the release of cytokines and host cell death. Here, we will focus on the most recent findings that have started to unravel the manifold restriction mechanism controlled by GBPs in mouse and human cells, and that shed light on the molecular cues that control GBP recruitment to bacterial membranes.

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“…Also, hGBP‐1 is able to guide both, hGBP‐2 and hGBP‐5, to its vesicle‐like structures within the cytosol . If those vesicle‐like structures represent hGBP‐1‐coated vesicles or the homomeric polymers that we identified recently remains elusive. Binding to membranes as well as polymer formation are abilities which are gained by the farnesyl moiety attached to hGBP‐1's CaaX box .…”

Section: Introductionmentioning

confidence: 91%

“…If those vesicle-like structures represent hGBP-1-coated vesicles or the homomeric polymers that we identified recently [29] remains elusive. Binding to membranes as well as polymer formation are abilities which are gained by the farnesyl moiety attached to hGBP-1's CaaX box [28][29][30][31]. However, the other biochemical properties of farnesylated hGBP-1 are not altered when compared with nonfarnesylated hGBP-1 [29].…”

Section: Introductionmentioning

confidence: 98%

Homo and hetero dimerisation of the human guanylate‐binding proteins hGBP‐1 and hGBP‐5 characterised by affinities and kinetics

2018

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The human guanylate-binding proteins (hGBPs) exhibit diverse antipathogenic and tumour-related functions which make them key players in the innate immune response. The isoforms hGBP-1 to hGBP-5 form homomeric complexes and localise to specific cellular compartments. Upon heteromeric interactions, hGBPs are able to guide each other to their specific compartments. Thus, homo- and heteromeric interactions allow the hGBPs to build a network within the cell which might be important for their diverse biological functions. We characterised homomeric complexes of hGBPs in vitro and presented most recently that nonprenylated hGBP-1 and hGBP-5 form dimers as highest oligomeric species while farnesylated hGBP-1 is able to form polymers. We continued to work on the biochemical characterisation of the heteromeric interactions between hGBPs and present here results for nonprenylated hGBP-1 and hGBP-5. Multiangle light scattering identified the GTP-dependent heteromeric complex as dimer. Also hGBP-5's tumour-associated splice variant (hGBP-5ta) was able to form a hetero dimer with hGBP-1. Intriguingly, both hGBP-5 splice variants were able to induce domain rearrangements within hGBP-1. We further characterised the homo and hetero dimers with Förster resonance energy transfer-based experiments. This allowed us to obtain affinities and kinetics of the homo and hetero dimer formation. Furthermore, we identified that the LG domains of hGBP-1 and hGBP-5 build an interaction site within the hetero dimer. Our in vitro study provides mechanistic insights into the homomeric and heteromeric interactions of hGBP-1 and hGBP-5 and present useful strategies to characterise the hGBP network further.

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Nucleotide-dependent farnesyl switch orchestrates polymerization and membrane binding of human guanylate-binding protein 1

Cited by 61 publications

References 56 publications

The human guanylate‐binding proteins hGBP‐1 and hGBP‐5 cycle between monomers and dimers only

The human guanylate‐binding proteins hGBP‐1 and hGBP‐5 cycle between monomers and dimers only

Sensing of invading pathogens by GBPs: At the crossroads between cell-autonomous and innate immunity

Homo and hetero dimerisation of the human guanylate‐binding proteins hGBP‐1 and hGBP‐5 characterised by affinities and kinetics

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