Formation of thyroid hormone revealed by a cryo-EM structure of native bovine thyroglobulin

Marechal, N.; Serrano, Banyuhay P.; Zhang, Xinyan; Weitz, Charles J.

doi:10.1038/s41467-022-30082-4

Cited by 11 publications

(8 citation statements)

References 22 publications

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“…[16] described the structure of non-deglycosylated TG from patients without known thyroid de ciencies and the atomic model of the dimeric TG covers about 90% of the molecule. At the same time, Kim et al [15] and Marechal et al [17] reported the structure of native bovine TG at an overall resolution of 2.6 Å and 3.3 Å, respectively. These models provide high resolution information on the structural organization of domains, formed disul de bridges and hormonogenic, proteolysis, and glycosylation sites.…”

Section: Discussionmentioning

confidence: 95%

“…The N-terminal and central region of the monomeric TG preprotein is composed of three different cysteine-rich repeat motifs called TG type 1, TG type 2 and TG type 3, whereas the región C-terminal is integrated by a non-repeat cholinesterase-like (ChEL) domain [8][9][10][11][12][13]. Recently, the 3-dimensional atomic structure of human an bovine TG has been reported [14][15][16][17] To date, more than two hundred and ninety deleterious variants in the human TG gene (loss-of-function variants or missense variants involving wild-type cysteine residues and the ChEL domain) have been reported associated to thyroid diseases, mainly to CH [18][19][20][21][22][23][24]. Several studies show that structurally defective TG mutants become trapped in the endoplasmic reticulum (ER), causing thyrocyte ER stress [25][26][27][28].…”

Section: Introductionmentioning

confidence: 99%

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The p.Pro2232Leu variant in the ChEL domain of thyroglobulin gene causes intracellular transport disorder and congenital hypothyroidism

et al. 2022

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Thyroglobulin (TG), the predominant glycoprotein of the thyroid gland, functions as matrix protein in thyroid hormonegenesis. TG de ciency results in thyroid dyshormonogenesis. These variants produce a heterogeneous spectrum of congenital goitre, with an autosomal recessive mode of inheritance. The purpose of this study was to identify and functionally characterize new variants in the TG gene in order to increase the understanding of the molecular mechanisms responsible for thyroid dyshormonogenesis. A total of four patients from two non-consanguineous families with marked alteration of TG synthesis were studied.The two families were previously analysed in our laboratory, only one deleterious allele, in each one, was detected after sequencing the TG gene (c.2359C > T [p.Arg787*], c.5560G > T [p.Glu1854*]). These ndings were con rmed in the present studies by Next-Generation Sequencing. The single nucleotide coding variants of the TG gene were then analyzed to predict the possible variant causing the disease. The p.Pro2232Leu (c.6695C > T), identi ed in both families, showing a low frequency population in gnomAD v2.1.1 database and protein homology, amino acid prediction, and 3D modeling analysis predict a potential pathogenic effect of this variant. We also transiently express p.Pro2232Leu in a full-length rat TG cDNA clone and con rmed that this point variant was su cient to cause intracellular retention of mutant TG in HEK293T cells. Consequently, each family carried a compound heterozygous for p.Arg787*/p.Pro2232Leu or p.Glu1854*/p.Pro2232Leu variants.In conclusion, our results con rm the pathophysiological importance of altered TG folding as a consequence of missense variants located in the ChEL domain of TG.

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Section: Discussionmentioning

confidence: 95%

Section: Introductionmentioning

confidence: 99%

The p.Pro2232Leu variant in the ChEL domain of thyroglobulin gene causes intracellular transport disorder and congenital hypothyroidism

et al. 2022

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“…Micrographs were processed with relion [35]. The streptavidin crystal pattern in the micrographs of the streptavidin affinity grids was erased in Fourier space using python scripts described in [36]. Particles were extracted with a binning of 4 times.…”

Section: Methodsmentioning

confidence: 99%

“…For the Rea1 ΔAAA2H2α ATPγS and Rea1 D2915A-R2976A-D3042A ATP data sets, standard copper/rhodium quantifoil R2/2 with an additional carbon layer and streptavidin affinity grids [36] were used. In addition, standard copper/rhodium quantifoil R2/2 without additional carbon layer were used for Rea1 D2915A-R2976A-D3042A ATP.…”

Section: Cryoem Electron Microscopymentioning

confidence: 99%

The Ribosome Maturation Factor Rea1 utilizes nucleotide independent and ATP-hydrolysis driven Linker remodelling for the removal of ribosome assembly factors

Busselez

Koenig

Klos

et al. 2022

Preprint

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The essential ribosome maturation factor Rea1 (also known as Midasin) catalyses the removal of assembly factors from precursors of the large ribosomal subunit and subsequently promotes their export from the nucleus to the cytosol. Rea1 is a large protein of nearly 5000 residues and a member of the AAA+ (ATPases associated with various cellular activities) protein family. It consists of a concatenated ring of six AAA+ domains from which the ≈ 1700 residue linker emerges that is subdivided into stem, middle and top domains. A flexible and unstructured D/E rich region connects the linker top to a MIDAS (metal ion dependent adhesion site) domain, which is able to bind the assembly factor substrates. Despite its key importance for ribosome maturation, the Rea1 mechanism driving assembly factor removal is still poorly understood. Although early studies on Rea1 demonstrated that assembly factor removal requires ATP hydrolysis, the Rea1 conformations associated with ATP hydrolysis have not been identified yet. Here we demonstrate that the Rea1 linker is essential for assembly factor removal. It rotates and swings towards the AAA+ ring following a complex remodelling scheme involving nucleotide independent as well as nucleotide dependent steps. ATP hydrolysis is required to engage the linker top with the AAA+ ring and ultimately with the AAA+ docked MIDAS domain. The interaction between the linker top and the MIDAS domain allows force transmission for assembly factor removal.

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“…The Tg protein concludes with the Cholinesterase-Like domain (ChEL, bearing three internal disulfide bonds) plus a short unique tail sequence ( Figure 1 A). Four recent papers have provided new insight into the three-dimensional structure of Tg (two human; two bovine) by modeling from cryogenic electron microscopy [ 28 , 34 , 40 , 41 ], and schematic of the 3D structure is shown in Figure 1 B. Specifically, when the color-coded image of Figure 1 B is considered alongside the color-coded primary structure of Tg in Figure 1 A, it becomes apparent that globally, the monomer structure has an overall J-like shape, in which region I is roughly equally divided into an ‘N-terminal domain’ encoded by the first 9 exons (of a total of 48), and what has been called the ‘core domain’ is encoded by the next 7 exons.…”

Section: Tg Folding and Tg Mutations That Trigger Misfoldingmentioning

confidence: 99%

Defective Thyroglobulin: Cell Biology of Disease

Zhang

Young

Morishita

et al. 2022

IJMS

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The primary functional units of the thyroid gland are follicles of various sizes comprised of a monolayer of epithelial cells (thyrocytes) surrounding an apical extracellular cavity known as the follicle lumen. In the normal thyroid gland, the follicle lumen is filled with secreted protein (referred to as colloid), comprised nearly exclusively of thyroglobulin with a half-life ranging from days to weeks. At the cellular boundary of the follicle lumen, secreted thyroglobulin becomes iodinated, resulting from the coordinated activities of enzymes localized to the thyrocyte apical plasma membrane. Thyroglobulin appearance in evolution is essentially synchronous with the appearance of the follicular architecture of the vertebrate thyroid gland. Thyroglobulin is the most highly expressed thyroid gene and represents the most abundantly expressed thyroid protein. Wildtype thyroglobulin protein is a large and complex glycoprotein that folds in the endoplasmic reticulum, leading to homodimerization and export via the classical secretory pathway to the follicle lumen. However, of the hundreds of human thyroglobulin genetic variants, most exhibit increased susceptibility to misfolding with defective export from the endoplasmic reticulum, triggering hypothyroidism as well as thyroidal endoplasmic reticulum stress. The human disease of hypothyroidism with defective thyroglobulin (either homozygous, or compound heterozygous) can be experimentally modeled in thyrocyte cell culture, or in whole animals, such as mice that are readily amenable to genetic manipulation. From a combination of approaches, it can be demonstrated that in the setting of thyroglobulin misfolding, thyrocytes under chronic continuous ER stress exhibit increased susceptibility to cell death, with interesting cell biological and pathophysiological consequences.

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Formation of thyroid hormone revealed by a cryo-EM structure of native bovine thyroglobulin

Cited by 11 publications

References 22 publications

The p.Pro2232Leu variant in the ChEL domain of thyroglobulin gene causes intracellular transport disorder and congenital hypothyroidism

The p.Pro2232Leu variant in the ChEL domain of thyroglobulin gene causes intracellular transport disorder and congenital hypothyroidism

The Ribosome Maturation Factor Rea1 utilizes nucleotide independent and ATP-hydrolysis driven Linker remodelling for the removal of ribosome assembly factors

Defective Thyroglobulin: Cell Biology of Disease

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