Extracellular signal-regulated kinases (ERKs) modulate cellular activities in response to extracellular stimuli and play important biological roles. Thus, perturbed kinase pathways induce pathological conditions, such as tumor development. Rit, a novel member of the Ras family GTPases, activase ERK6, and its over-expression confers tumorigenicity. We hypothesized the presence of scaffolding molecules specific to ERK6, similar to other known MAP kinases. We performed yeast two-hybrid assays using ERK6 as bait, and Scribble was identified as a binding partner. Scribble contains 16 LRR domains and four PDZ domains. We performed immunoprecipitation (IP) assays and discovered ERK2 as another binding partner. Surprisingly, no interaction was observed with the highly homologous MAP kinase, ERK1. No other representative kinases showed binding capabilities with Scribble. IP data confirmed that both ERK2 and ERK6 bind to Scribble through its LRR and PDZ domains. Deletion of ten aminoi acids from the C-terminus of ERK2 and ERK6 abolished these interactions. In vitro kinase assays indicated the kinase suppressing ability of Scribble. Focus formation assays were performed with RitQ79L and H-RasV12 as constitutive activators of ERK6 and ERK2, respectively, in the presence of Scribble. Results confirmed the role of Scribble as a tumor suppressor.

Mammalian Scribble is a target for proteins encoded by human papilloma virus, retro- and flaviviruses. Tick-borne encephalitis virus (TBEV) is a flavivirus that have evolved distinct strategies to escape antiviral responses. Information of how flaviviruses intrude on cell integrity comes from understanding of the roles that host-factors play when they interfere with viruses. The first part of this thesis describes a novel interaction between the TBEVNS5 protein and Scribble. The importance of the interaction was demonstrated by RNAi-mediated depletion of Scribble, which prevented suppression of JAK-STAT signaling by NS5. Together, these results define Scribble as a novel target for NS5. TBEV is known to cause central nervous system disease TBE in humans that can lead to cognitive dysfunction. A unifying theme in CNS related diseases are defects in neuronal extensions. We therefore addressed the effects of TBEV expression in PC12 cell differentiation, which is characterized by extensive neurite growth. Our data show that TBEVNS5 suppresses neurite outgrowth through the Rho GTPase Rac1. These findings provide evidence that Rac1 is an indirect target of NS5 in neurite inhibition. Scribble was recently implicated in spine morphogenesis. Thus, we tested the role of Scribble in neurite elongation. Depletion of Scribble in PC12 cells, reduced neurite density but increased length of those remaining. Moreover, Scribble bound components in the Ras/ERK cascade in a growth factor dependent manner. Together, these results demonstrate that Scribble controls neurite elongation by scaffolding MAPK components. Moreover, as loss of dendritic spines, actin-rich protrusions on neurons, is a feature in cognitive dysfunction we speculate that cognitive dysfunction in TBE might involve disturbed Scribble expression by NS5. We also investigated the binding between NS1 of Influenza A virus and Scribble. The PDZ domains of Scribble are usually selective for specific C-terminal motifs in proteins. Because NS1 has a canonical PDZ motif we tested if binding to Scribble depends on this motif. We found that Scribble binds NS1; the association is dependent on the NS1 C-terminus that is recognized by PDZ3-4 of Scribble. Together, these results suggest that Scribble is a target for the H5N1 NS1 protein / At the time of the doctoral defense, the following papers were unpublished and had a status as follows: Paper 2: In press. Paper 3: Manuscript. Paper 4: Manuscript.

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The main purpose of this thesis is to explore experiences of two different psychological interventions based on art psychotherapy in women with a psychological or physical illness. The two interventions are art psychotherapy and art therapy. The difference between these two interventions is that the art therapist works with the transference in art psychotherapy but not in art therapy. The thesis consists of two studies of art psychotherapy: An art psychotherapy intervention is evaluated in Study 1 (papers III and V) which examines a group of patients diagnosed with depression and Study 2 (paper II) which examines experiences in a group of six patients diagnosed with vulva vestibulitis. An art therapy intervention is evaluated in the third study (papers I and IV); where experiences in patients diagnosed with breast cancer are examined. In Study 1, forty-three (n=43) depressed women were randomly assigned to either an intervention group or a control group (verbal psychotherapy). The aim was to examine the outcome of time limited psychodynamic art therapy compared to time-limited psychodynamic verbal therapy for patients with depressive symptoms. Interviews were performed before, immediately after, and three months after the termination of psychotherapy, and self-rating scales which focus on stress reactions, depression and symptoms as well as an observer rating scale on depression were used. The interviews and the art sessions were video-recorded, and the verbal psychotherapy was tape-recorded. The results showed that the art and verbal psychotherapies were comparable. The conclusion was that short-term psychodynamic art psychotherapy could be a valuable treatment for depressed women. In an in-depth content analysis, the method of scribbling was further investigated and exemplified with the therapies of two participants. In this study, the patients’ pictures and verbal expressions of progress, along with considerations of how to interpret the pictures were in focus. When leaving therapy the two patients took advantage of the paper, made complete forms, symbolised in words what they have expressed in pictures; in pace with psychotherapy the themes alter towards separation, individuation, and attempt to relate in a new way. The conclusion was that limelimited psychodynamic art therapy suggests giving a safer place for the self as the cohesion is firmer with better boundaries. Study 2 is a pilot study, which involved six young patients newly diagnosed with vulva vestibulitis. The aim of the study was to investigate pain at vestibulum, mental health, and self-image after fifteen sessions of art psychotherapy. Five of the patients were judged to have less pain three months after termination of therapy. The conclusion was that art psychotherapy with its openness seemed to affect young women in their experiences of vulva vestibulitis in a positive direction. Study 3 examined the potential benefit of art therapy for women with primary breast cancer. The sample comprised forty-one (n=41) patients who were randomly assigned either to an art therapy group or to a control group. The art therapy was going on during five weeks radiation treatment, one session per week. The aim was to investigate the outcome of art therapy, to quantify and compare the participant coping s, self-image, and the symptoms with the participant in the control group. Interviews were performed before, immediately after, and six month after inclusion. A set of self-rating scales was used: Coping Resources Inventory, the Structural Analysis of Social Behavior, and Symptom Check List – 90. The result showed that the patients in the art therapy group rated their coping s and especially their social s, higher than the control group, and that the average patients in the art therapy group improved in depressive symptoms and symptoms of anxiety, and that the general psychiatric symptoms improved as well. A linear regression analysis showed a tendency that the coping s increased in the art therapy group and decreased in the control group or even stagnated in the social domain. A second report on self-image, symptoms, treatment, and social variables showed that art therapy was related to lower ratings of depression, anxiety, and general symptoms after treatment; chemotherapeutic treatment predicted lower depressive symptoms and general symptoms in contrast to axilliary surgery and hormonal treatment. The results showed that art therapy could be valuable complementary therapy in routine oncology practise. The conclusion is that art therapy can have a positive long-term effect on the crisis following the primary breast cancer and its consequences. Conclusion: The results show that time-limited psychodynamic art psychotherapy is valuable for depressed women; that it is a valuable complement for women with vulva vestibulitis; and that art therapy is a valuable complement in the care and cure of women with primary breast cancer.

La voie de signalisation Ras/MAPK régule de nombreuses fonctions biologiques et occupe un rôle central dans la transmission de signaux extracellulaires à des protéines cibles intracellulaires. Les dysfonctionnements de cette voie sont responsables de plusieurs maladies et syndromes génétiques, tels que le cancer ou le diabète. Cette voie de signalisation, qui régule l’activité des protéines kinases ERK1/2 comptant un grand nombre de substrats cellulaires, occupe une place primordiale dans de nombreux processus biologiques. Parmi ces substrats, on retrouve les protéines kinases de la famille RSK qui comptent quatre isoformes (RSK1-4). Bien que plusieurs substrats cellulaires aient été identifiés pour les isoformes RSK1 et RSK2, les fonctions biologiques des kinases RSK ainsi que les mécanismes moléculaires les régulant sont encore aujourd’hui peu décrits. Ainsi, afin d’améliorer nos connaissances sur la famille des RSK, nous avons utilisé plusieurs approches. Tout d’abord, nous avons déterminé les partenaires cellulaires à proximité des kinases RSK avec la mise en place d’une méthode protéomique spécifique. Cette première étape nous a permis d’identifier la protéine p120ctn comme un nouveau substrat des kinases RSK, mais aussi de démontrer le rôle de ces dernières dans la régulation des jonctions intercellulaires. D’autre part, en se focalisant sur un domaine particulier des kinases RSK encore non étudié, notre deuxième étude apporte elle aussi de nouvelles connaissances sur les différentes interactions des protéines RSK. Ces travaux ont entre autres permis de montrer que la liaison de l’isoforme RSK2 avec la protéine Scribble inhibe son activation par la voie de signalisation Ras/MAPK. En établissant donc des études à grande échelle pour déterminer les interactions propres à chaque isoforme des kinases RSK, nous avons identifié plusieurs nouveaux partenaires cellulaires de ces protéines ainsi que leurs fonctions associées. Cette étape est cruciale à la compréhension et la caractérisation du rôle des protéines RSK, notamment dans le développement des cellules cancéreuses. / The Ras/MAPK signaling pathway regulates many biological functions and plays a key role in transducing extracellular signals to intracellular target proteins. Inappropriate regulation of this pathway leads to a variety of diseases and genetic syndromes, including cancer or diabetes. This signaling pathway regulates the activity of ERK1/2 protein kinases, which have many cellular substrates, and therefore regulates significant biological processes. Among these substrates, there is the RSK (p90 ribosomal S6 kinase) family of protein kinases, which is composed of four isoforms (RSK1-4). Although several cellular substrates have been identified for the RSK1 and RSK2 isoforms, the biological functions of RSK kinases and the molecular mechanisms regulating them are still poorly understood. Thus, to improve our knowledge of the RSK family, we used several approaches. First, we determined the cellular partners of the RSK kinases using a proximity-based labeling technique. This first step allowed us to identify the p120ctn protein as a new substrate of RSK kinases, but also to demonstrate the role of these proteins in the regulation of intercellular junction’s integrity. Additionally, by focusing on a particular domain of RSK kinases still unstudied, our second study also provided new insights into the different interactions of RSK proteins. Finally, we demonstrated that the binding of the RSK2 isoform with the Scribble protein inhibits its activation by the Ras/MAPK signaling pathway. Consequently, by establishing large-scale studies to determine the specific interactions of each RSK isoform, we have identified several new cellular partners of these proteins and their associated functions. This step is crucial to understand and characterize the role of the RSK proteins, particularly with respect to their described functions in cancer.