Decoding the PTM-switchboard of Notch

Antfolk, Daniel; Antila, Christian; Kemppainen, Kati; Landor, Sebastian K.-J.; Sahlgren, Cecilia

doi:10.1016/j.bbamcr.2019.07.002

Cited by 36 publications

(36 citation statements)

References 284 publications

(194 reference statements)

Supporting

Mentioning

Contrasting

Order By: Relevance

“…Since most proteins are phosphorylated, many on multiple sites 53 , the characterization of phosphorylation has become a paramount issue for understanding the fine-tuning of protein expression and activity. One notable example is represented by the Notch receptor's family 54 .…”

Section: Proline-rich Proteinmentioning

confidence: 99%

“…They are single-pass transmembrane proteins that share a characteristic three-domain structure: an extracellular domain (NECD), a transmembrane region (NTM) and an intracellular domain (NICD), which is able to translocate into the nucleus and activate transcription of target genes 55 . NICD activity is fine-tuned 56 , for instance by dynamic PTMs 54 . However, little is known about the phosphorylation of the intracellular domain of Notch3 paralog.…”

Section: Proline-rich Proteinmentioning

confidence: 99%

See 1 more Smart Citation

ProAlanase is an Effective Alternative to Trypsin for Proteomics Applications and Disulfide Bond Mapping

Samodova

Hosfield

Cramer

et al. 2020

Molecular & Cellular Proteomics

View full text Add to dashboard Cite

Trypsin is the protease of choice in bottom-up proteomics. However, its application can be limited by the amino acid composition of target proteins and the pH of the digestion solution. In this study we characterize ProAlanase, a protease from the fungus Aspergillus niger that cleaves primarily on the C-terminal side of proline and alanine residues. ProAlanase achieves high proteolytic activity and specificity when digestion is carried out at acidic pH (1.5) for relatively short (2 h) time periods. To elucidate the potential of ProAlanase in proteomics applications, we conducted a series of investigations comprising comparative multi-enzymatic profiling of a human cell line proteome, histone PTM analysis, ancient bone protein identification, phosphosite mapping and de novo sequencing of a proline-rich protein and disulfide bond mapping in monoclonal antibody. The results demonstrate that ProAlanase is highly suitable for proteomics analysis of the arginine- and lysine-rich histones, enabling high sequence coverage of multiple histone family members. It also facilitates an efficient digestion of bone collagen thanks to the cleavage at the C-terminus of hydroxyproline which is highly prevalent in collagen. This allows to identify complementary proteins in ProAlanase- and trypsin-digested ancient bone samples, as well as to increase sequence coverage of non-collagenous proteins. Moreover, digestion with ProAlanase improves protein sequence coverage and phosphosite localization for the proline-rich protein Notch3 intracellular domain (N3ICD). Furthermore, we achieve a nearly complete coverage of N3ICD protein by de novo sequencing using the combination of ProAlanase and tryptic peptides. Finally, we demonstrate that ProAlanase is efficient in disulfide bond mapping, showing high coverage of disulfide-containing regions in a non-reduced monoclonal antibody.

show abstract

Section: Proline-rich Proteinmentioning

confidence: 99%

Section: Proline-rich Proteinmentioning

confidence: 99%

ProAlanase is an Effective Alternative to Trypsin for Proteomics Applications and Disulfide Bond Mapping

Samodova

Hosfield

Cramer

et al. 2020

Molecular & Cellular Proteomics

View full text Add to dashboard Cite

show abstract

“…Transcription factor in concert with chromatin modifiers establish a chromatin environment that predetermines specificity of the Notch target gene expression (3)(4)(5). In regard to posttranslational modifications (PTMs), not only histone tails, but also the Notch coactivator complex are known to be phosphorylated, methylated, acetylated, SUMOylated and ubiquitinylated (6)(7)(8). These modifications control not only the stability of Notch but also determine amplitude and duration of the Notch response (9) and can be incorporated upon cellular stress signals (10).…”

Section: Introductionmentioning

confidence: 99%

SUMOylated Non-canonical Polycomb PRC1.6 Complex as a Prerequisite for Recruitment of Transcription Factor RBPJ

Sotomska

Liefke

Ferrante

et al. 2021

Preprint

View full text Add to dashboard Cite

BackgroundNotch signaling controls cell fate decisions in many contexts during development and adult stem cell homeostasis and, when dysregulated, leads to carcinogenesis. The central transcription factor RBPJ assembles the Notch coactivator complex in the presence of Notch signalling, and represses Notch target gene expression in its absence.ResultsWe identified L3MBTL2 and additional members of the non-canonical polycomb repressive PRC1.6 complex in DNA-bound RBPJ associated complexes and demonstrate that L3MBTL2 directly interacts with RBPJ. Depletion of RBPJ does not affect occupancy of PRC1.6 components at Notch target genes. Conversely, absence of L3MBTL2 reduces RBPJ occupancy at enhancers of Notch target genes. Since L3MBTL2 and additional members of the PRC1.6 are known to be SUMOylated, we investigated whether RBPJ uses SUMO-moieties as contact points. Indeed, we found that RBPJ binds to SUMO2/3 and that this interaction depends on a defined SUMO-interaction motif. Furthermore, we show that pharmacological inhibition of SUMOylation reduces RBPJ occupancy at Notch target genes.ConclusionsWe propose that the PRC1.6 complex and its conjugated SUMO-modifications provide a scaffold that is recognized by RBPJ and promotes its recruitment to Notch target genes.

show abstract

“…Beyond these large domains are several smaller structural features and a wide array of post-translational modifications. These modifications range from the assortment of glycosylations on the EGF-like repeats, to protein attachments (ubiquitination/sumoylation), and a plethora of other transient, amino-acid level modifications (phosphorylations/acetylations/methylations/hydroxylations), all thoroughly reviewed in Antfolk et al, (2019) 57 . Other minor, yet critical, structural features include two nuclear localization signals (NLS) 50,58 and a Proline/Glutamic acid/Serine/Threonine (PEST) domain, which is involved in the Notch protein's rapid degradation 59,60 .…”

Section: The Intracellular Domainmentioning

confidence: 99%

Dimerization of the Notch Intracellular Domain Results in Distinct Signaling Activity

Crow¹

View full text Add to dashboard Cite

The Notch signaling pathway is a core component of multicellularity; enabling cells to directly communicate with both their neighbors and the surrounding microenvironment. These signals are translated directly through the Notch proteins, where a fragment of Notch transitions into the nucleus to act as a co-transcription factor, setting into motion a host of physiological responses. Commonly involved in pathways that define a cell’s identity and fate decisions, what appears to be a simplistic pathway instead exists in a state of high-tunability and strict control. Missteps in this pathway are generally embryonically lethal or lead to a suite of congenital disorders and cancers. Therefore, it’s pertinent to understand the mechanisms of Notch that provide its flexibility and pleiotropic outcomes. One such property is its ability to homodimerize on DNA while within its transcriptional activation complex, resulting in an enhanced transcriptional signal of a select pool of Notch target genes. This dissertation reviews the general mechanics behind Notch signaling, discusses how the field of Notch dimerization came to be and where it stands currently, and finally, details my contributions to the understanding of this regulatory mechanism. Despite Notch’s ubiquitous function in metazoan life, there are still many mysteries behind this signaling pathway. The work detailed here describes my time spent as a basic science researcher, where my findings contribute a couple of puzzle pieces to the expansive Notch signaling field.

show abstract

Decoding the PTM-switchboard of Notch

Cited by 36 publications

References 284 publications

ProAlanase is an Effective Alternative to Trypsin for Proteomics Applications and Disulfide Bond Mapping

ProAlanase is an Effective Alternative to Trypsin for Proteomics Applications and Disulfide Bond Mapping

SUMOylated Non-canonical Polycomb PRC1.6 Complex as a Prerequisite for Recruitment of Transcription Factor RBPJ

Dimerization of the Notch Intracellular Domain Results in Distinct Signaling Activity

Contact Info

Product

Resources

About