Differential splicing of the lectin domain of an O-glycosyltransferase modulates both peptide and glycopeptide preferences

May, Carolyn; Ji, Suena; Syed, Zulfeqhar A.; Revoredo, Leslie; Daniel, Earnest James Paul; Gerken, Thomas A.; Tabak, Lawrence A.; Samara, N.L.; Hagen, Karl S.

doi:10.1074/jbc.ra120.014700

Cited by 7 publications

(5 citation statements)

References 46 publications

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“…36,37 The charge state surrounding T or S may be a factor because the polarity of the GalNAcTs lectin domain affects glycosylation. 38 According to N-glycopeptide analysis, spike S1 also showed different O-glycosylation expressed in HEK293 and baculovirus-insect cells. In the HEK293 cells, T323 and T325 are Oglycosylated by GalNAc and GalGalNAc mucin-type Oglycans.…”

Section: ■ Results and Discussionmentioning

confidence: 99%

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Impact of Expressing Cells on Glycosylation and Glycan of the SARS-CoV-2 Spike Glycoprotein

Wang

et al. 2021

ACS Omega

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The spike glycoprotein of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2), the first point of contact for the virus to recognize and bind to host receptors, is the focus of biomedical research seeking to effectively prevent and treat coronavirus disease (COVID-19). The mass production of spike glycoproteins is usually carried out in different cell systems. Studies have been shown that different expression cell systems alter protein glycosylation of hemagglutinin and neuraminidase in the influenza virus. However, it is not clear whether the cellular system affects the spike protein glycosylation. In this work, we investigated the effect of an expression system on the glycosylation of the spike glycoprotein and its receptor-binding domain. We found that there are significant differences in the glycosylation and glycans attached at each glycosite of the spike glycoprotein obtained from different expression cells. Since glycosylation at the binding site and adjacent amino acids affects the interaction between the spike glycoprotein and the host cell receptor, we recognize that caution should be taken when selecting an expression system to develop inhibitors, antibodies, and vaccines.

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Section: ■ Results and Discussionmentioning

confidence: 99%

“…E.C. 2.4.1.41)) accessible to the T or S residues. , The charge state surrounding T or S may be a factor because the polarity of the GalNAcTs lectin domain affects glycosylation …”

Section: Resultsmentioning

confidence: 99%

Impact of Expressing Cells on Glycosylation and Glycan of the SARS-CoV-2 Spike Glycoprotein

Wang

et al. 2021

ACS Omega

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“…If so, they are novel. These include Cbp53E , an ortholog of calbindin known to affect axon branching in Drosophila [ 103 ], and pgant9 , an enzyme involved in the sugar-modification of proteins [ 104 , 105 ]. While further validation will be needed, we suggest that concordance across datasets may justify further investigation of these and other DEGs.…”

Section: Discussionmentioning

confidence: 99%

Transcriptional changes in specific subsets of Drosophila neurons following inhibition of the serotonin transporter

Bonanno

Krantz

2023

Transl Psychiatry

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The transcriptional effects of SSRIs and other serotonergic drugs remain unclear, in part due to the heterogeneity of postsynaptic cells, which may respond differently to changes in serotonergic signaling. Relatively simple model systems such as Drosophila afford more tractable microcircuits in which to investigate these changes in specific cell types. Here, we focus on the mushroom body, an insect brain structure heavily innervated by serotonin and comprised of multiple different but related subtypes of Kenyon cells. We use fluorescence-activated cell sorting of Kenyon cells, followed by either bulk or single-cell RNA sequencing to explore the transcriptomic response of these cells to SERT inhibition. We compared the effects of two different Drosophila Serotonin Transporter (dSERT) mutant alleles as well as feeding the SSRI citalopram to adult flies. We find that the genetic architecture associated with one of the mutants contributed to significant artefactual changes in expression. Comparison of differential expression caused by loss of SERT during development versus aged, adult flies, suggests that changes in serotonergic signaling may have relatively stronger effects during development, consistent with behavioral studies in mice. Overall, our experiments revealed limited transcriptomic changes in Kenyon cells, but suggest that different subtypes may respond differently to SERT loss-of-function. Further work exploring the effects of SERT loss-of-function in other circuits may be used help to elucidate how SSRIs differentially affect a variety of different neuronal subtypes both during development and in adults.

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“…[99], and pgant9, an enzyme involved in sugar-modification of proteins[100,101]. While further validation will be needed, we suggest that concordance across some datasets may justify…”

mentioning

confidence: 83%

Transcriptional changes in specific subsets of Drosophila neurons following inhibition of the serotonin transporter

Krantz

Bonanno

2023

Preprint

View full text Add to dashboard Cite

The transcriptional effects of SSRIs and other serotonergic drugs remain unclear, in part due to the heterogeneity of postsynaptic cells, which may respond differently to changes in serotonergic signaling. Relatively simple model systems such as Drosophila afford more tractable microcircuits in which to investigate these changes in specific cell types. Here, we focus on the mushroom body, an insect brain structure heavily innervated by serotonin and comprised of multiple different but related subtypes of Kenyon cells. We use fluorescence activated cell sorting of Kenyon cells, followed by either or bulk or single cell RNA sequencing to explore the transcriptomic response of these cells to SERT inhibition. We compared the effects of two different Drosophila Serotonin Transporter (dSERT) mutant alleles as well as feeding the SSRI citalapram to adult flies. We find that the genetic architecture associated with one of the mutants contributed to significant artefactual changes in expression. Comparison of differential expression caused by loss of SERT during development versus aged, adult flies, suggests that changes in serotonergic signaling may have relatively stronger effects during development, consistent with behavioral studies in mice. Overall, our experiments revealed limited transcriptomic changes in Kenyon cells, but suggest that different subtypes may respond differently to SERT loss-of-function. Further work exploring the effects of SERT loss-of-function in other Drosophila circuits may be used help to elucidate how SSRIs differentially affect a variety of different neuronal subtypes both during development and in adults.

show abstract

Differential splicing of the lectin domain of an O-glycosyltransferase modulates both peptide and glycopeptide preferences

Cited by 7 publications

References 46 publications

Impact of Expressing Cells on Glycosylation and Glycan of the SARS-CoV-2 Spike Glycoprotein

Impact of Expressing Cells on Glycosylation and Glycan of the SARS-CoV-2 Spike Glycoprotein

Transcriptional changes in specific subsets of Drosophila neurons following inhibition of the serotonin transporter

Transcriptional changes in specific subsets of Drosophila neurons following inhibition of the serotonin transporter

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