Pawel Borowicz

• Borowicz, Pawel; Chan, Hanna; Hauge, Anette & Spurkland, Anne (2020). Adaptor proteins: Flexible and dynamic modulators of immune cell signalling. Scandinavian Journal of Immunology.  ISSN 0300-9475.  92(6) . doi: 10.1111/sji.12951 Show summary

  To maintain homeostasis, all cells respond to environmental cues via a multitude of surface receptors. In order to act appropriately in their environment, cells are dependent on the transduction of the incoming signal through tightly regulated and interconnected signalling pathways to the cell nucleus. In particular, cells implicated in the immune system greatly depend on such systems to respond in a flexible and dynamic manner to environmental challenges. One major group of intracellular proteins that are involved in these signalling pathways are adaptor proteins. Although adaptor proteins are essential for normal immune cell operation, the functional role of this group of signalling proteins remains to be fully appreciated. So far, research on adaptor proteins has revealed their unique potential in building transient complexes in a reversible, dynamic and inducible manner. In this review, we explore the roles of adaptor proteins – in space and time of intracellular signalling – and their associations with human disease. Examples of adaptor proteins expressed in hematopoietic cells highlight their crucial role in the immune system. Lastly, we present challenges faced in elucidating roles of adaptor proteins, as illustrated by the T cell–specific adaptor (TSAd) protein encoded by the SH2D2A gene.

• Borowicz, Pawel; Chan, Hanna; Talmo Medina, Daniel; Gumpelmair, Simon; Kjelstrup, Hanna & Spurkland, Anne (2019). A simple and efficient workflow for generation of knock-in mutations in Jurkat T cells using CRISPR/Cas9. Scandinavian Journal of Immunology.  ISSN 0300-9475. . doi: 10.1111/sji.12862 Full text in Research Archive.

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• Borowicz, Pawel & Spurkland, Anne (2020). Regulation of T cell activation by two conserved phosphotyrosines in Lck and its adapter protein TSAd. Full text in Research Archive. Show summary

  T cells are major players in the immune system. These cells can recognize dangerous antigens through their specific receptors. Harnessing the power of T cells would be beneficial for not only fighting numerous infectious diseases, but also for treating cancer or preventing autoimmune diseases. However, the intracellular signalling involved in the T-cell receptor (TCR) pathway is remarkably complex. It is necessary to understand it more fully in order to control the T cells behaviour. We focused on two proteins involved in TCR signalling. First, lymphocyte-specific protein tyrosine kinase (Lck) starts a phosphorylation cascade downstream of TCR stimulation. Lck itself is tightly controlled by the phosphorylation of two tyrosines: Tyr394 and Tyr505. Recently a third tyrosine, Tyr192, emerged as an important regulatory site. Second, T cell specific adaptor protein (TSAd) is best known for its interaction with Lck. Its main function is to facilitate binding between other proteins. TSAd itself contains a number of phosphotyrosines. One of the most interesting is Tyr290, as it is highly conserved, but it is not predicted to be a prototypic binding site. The aim of this thesis was to understand the function of two phosphotyrosines, Tyr192 in Lck and Tyr290 in TSAd, in order to better comprehend the intracellular signalling of the TCR. The study was based on the knock-in cell lines generated with CRISPR/Cas9 genome editing. The mutated cell lines were studied using a wide spectrum of molecular biology techniques, including mass spectrometry, western blotting, immunoprecipitation and flow cytometry. Our findings revealed that Lck Tyr192 could control and stabilize Lck activity, both positively and negatively. TSAd Tyr290 could also control the protein’s activity. Our results suggest that this tyrosine most likely facilitates the proteins conformation. We have shown that both phosphotyrosines could influence the whole TCR signalling pathway.

• Borowicz, Pawel; Gopalakrishnan, Ramakrishna Prabhu; Chan, Hanna; Kjelstrup, Hanna; Foss, Stian; De Souza, Gustavo Antonio; Sundvold-Gjerstad, Vibeke & Spurkland, Anne (2019). A conserved pTyr motif in the unstructured C-terminus regulates the function of the T cell specific adaptor TSAd.
• Borowicz, Pawel; Sundvold-Gjerstad, Vibeke; Chan, Hanna; Abrahamsen, Greger; Kjelstrup, Hanna; Medina, Daniel; Nyman, Tuula Anneli & Spurkland, Anne (2019). Tyr192 in the Lck SH2 domain affects the regulation of Lck activity.
• Borowicz, Pawel; Sundvold-Gjerstad, Vibeke; Abrahamsen, Greger; Kjelstrup, Hanna; Nyman, Tuula Anneli & Spurkland, Anne (2018). Lck’s interactome is influenced by phosphorylation of Tyr192 in the Lck SH2 domain.
• Borowicz, Pawel; Sundvold-Gjerstad, Vibeke; Abrahamsen, Greger; Kjelstrup, Hanna; Nyman, Tuula Anneli & Spurkland, Anne (2018). Lck’s interactome is influenced by phosphorylation of Tyr192 in the Lck SH2 domain.
• Borowicz, Pawel; Sundvold-Gjerstad, Vibeke; Abrahamsen, Greger; Kjelstrup, Hanna; Nyman, Tuula Anneli & Spurkland, Anne (2018). Lck’s interactome is influenced by phosphorylation of Tyr192 in the Lck SH2 domain.
• Borowicz, Pawel; Gopalakrishnan, Ramakrishna Prabhu; De Souza, Gustavo Antonio; Abrahamsen, Greger; Sundvold-Gjerstad, Vibeke & Spurkland, Anne (2017). The interactome of TSAd phosphopeptides in T cells.
• Borowicz, Pawel; Sundvold-Gjerstad, Vibeke; Abrahamsen, Greger; Kjelstrup, Hanna & Spurkland, Anne (2017). The regulatory role of Lck Tyr192 in T cell receptor signalling.
• Borowicz, Pawel & Spurkland, Anne (2016). Intracellular signaling in CRISPR mutated Jurkat T cells.
• Borowicz, Pawel; Sundvold-Gjerstad, Vibeke; Abrahamsen, Greger & Spurkland, Anne (2016). Generation of CRISPR/Cas9 mutants in order to study TCR signalling pathway.

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