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Stevenson, N. L.

Publications and source records attributed to Stevenson, N. L..

3 recordsLinked to original sources

Morphological determinants of glycosylation efficiency in Golgi cisternae

The Golgi apparatus has an intricate spatial structure characterized by flattened membrane-bound compartments, known as cisternae. Cisternae house integral membrane enzymes that catalyse glycosylation, the addition of polymeric sugars to protein cargo, which is important for the trafficking and function of the products. The unusual and specific shape of Golgi cisternae is highly conserved across eukaryotic cells, suggesting significant influence in the correct functioning of the Golgi. Motivated by experimental evidence that disruption to Golgi morphology can lead to observable changes in secreted cargo mass distribution, we develop and analyse a mathematical model of polymerisation in a cisterna that combines chemical kinetics, spatial diffusion and adsorption and desorption between lumen and membrane. Exploiting the slender geometry, we derive a non-local nonlinear advection-diffusion equation that predicts secreted cargo mass distribution as a function of cisternal shape. The model predicts a maximum cisternal thickness for which successful glycosylation is possible, demonstrates the existence of an optimal thickness for most efficient glycosylation, and suggests how kinetic and geometric factors may combine to promote or disrupt polymer production. Author SummaryThe Golgi apparatus is a universal feature of eukaryotic cells, playing a critical role in post-translational modification of secreted proteins. Its importance is demonstrated by the variety of disorders caused by errors in its function. A major post-translational modification is glycosylation: the addition of sugar chains to protein cargo. The Golgi has a distinctive structure, being comprised of stacks of thin, hollow, flattened, cisternae. We develop a mathematical model of glycosylation, combining enzyme-mediated membrane-bound reactions with intra-cisternal diffusion, to quantify the effect of cisternal morphology on processing rate. The model identifies optimal and maximal cisternal thicknesses for glycosylation to proceed, in terms of biochemical parameters. The model offers a quantitative connection between the Golgis cisternal morphology and its function.

biophysics↗

Dynein-2 is required for a functional transition zone and bidirectional ciliary trafficking

The dynein-2 microtubule motor is the retrograde motor for intraflagellar transport. Mutations in dynein-2 components cause skeletal ciliopathies, notably Jeune syndrome. Dynein-2 comprises a heterodimer of two non-identical intermediate chains, WDR34 and WDR60. Here, we use knockout cell lines to demonstrate that each intermediate chain has a distinct role in cilia function. Both proteins are required to maintain a functional transition zone and for efficient bidirectional intraflagellar transport, only WDR34 is essential for axoneme extension. In contrast, only WDR60 is essential for co-assembly of the other subunits. Furthermore, WDR60 cannot compensate for loss of WDR34 or vice versa. This work defines a functional asymmetry to match the subunit asymmetry within the dynein-2 motor. Analysis of causative point mutations in WDR34 and WDR60 can partially restore function to knockout cells. Our data show that Jeune syndrome is caused by defects in transition zone architecture as well as intraflagellar transport.\n\nSUMMARYHere, Vuolo and colleagues use engineered knockout human cell lines to define roles for dynein-2 intermediate chains. WDR34 is required for axoneme extension, while WDR60 is not. Both subunits are required for cilia transition zone organization and bidirectional intraflagellar transport.

cell biology↗

The Golgi matrix protein giantin is required for normal cilia function in zebrafish

The Golgi is essential for glycosylation of newly synthesised proteins including almost all cell-surface and extracellular matrix proteoglycans. Giantin, encoded by the golgb1 gene, is a member of the golgin family of proteins that reside within the Golgi stack but its function remains elusive. Loss-of-function of giantin in rats causes osteochondrodysplasia; knockout mice show milder defects, notably a cleft palate. In vitro, giantin has been implicated in Golgi organization, biosynthetic trafficking, and ciliogenesis. Here we show that loss-of-function of giantin in zebrafish, using either morpholino or knockout techniques, causes defects in cilia function. Giantin morphants have fewer cilia in the neural tube and those remaining are longer. Mutants have the same number of cilia in the neural tube but these cilia are also elongated. Scanning electron microscopy shows that loss of giantin results in an accumulation of material at the ciliary tip, consistent with a loss-of-function of retrograde intraflagellar transport. Mutants show milder defects than morphants consistent with adaptation to loss of giantin.\n\nSummary statementLoss of giantin following either morpholino injection or genome editing in zebrafish results in defects in ciliogenesis.

developmental biology↗