umu.sePublications
Change search
CiteExportLink to record
Permanent link

Direct link
Cite
Citation style
  • apa
  • ieee
  • modern-language-association-8th-edition
  • vancouver
  • Other style
More styles
Language
  • de-DE
  • en-GB
  • en-US
  • fi-FI
  • nn-NO
  • nn-NB
  • sv-SE
  • Other locale
More languages
Output format
  • html
  • text
  • asciidoc
  • rtf
Novel mechanisms of ALK activation revealed by analysis of the Y1278S neuroblastoma mutation
Institute of Biomedicine,Dept of Medical Biochem and Cell Biol, Sahlgrenska Academy, University of Gothenburg.. (Professor Bengt Hallberg)
Institute of Biomedicine,Dept of Medical Biochem and Cell Biol, Sahlgrenska Academy, University of Gothenburg.. (Professor Ruth H Palmer)
Institute of Biomedicine,Dept of Medical Biochem and Cell Biol, Sahlgrenska Academy, University of Gothenburg.. (Professor Bengt Hallberg)
Institute of Biomedicine,Dept of Medical Biochem and Cell Biol, Sahlgrenska Academy, University of Gothenburg.. (Professor Ruth H Palmer)
Show others and affiliations
2017 (English)In: Cancers, ISSN 2072-6694, Vol. 9, no 11, article id 149Article in journal (Refereed) Published
Abstract [en]

Numerous mutations have been observed in the Anaplastic Lymphoma Kinase (ALK) receptor tyrosine kinase (RTK) in both germline and sporadic neuroblastoma. Here, we have investigated the Y1278S mutation, observed in four patient cases, and its potential importance in the activation of the full length ALK receptor. Y1278S is located in the 1278-YRASYY-1283 motif of the ALK activation loop, which has previously been reported to be important in the activation of the ALK kinase domain. In this study, we have characterized activation loop mutations within the context of the full length ALK employing cell culture and Drosophila melanogaster model systems. Our results show that the Y1278S mutant observed in patients with neuroblastoma harbors gain-of-function activity. Secondly, we show that the suggested interaction between Y1278 and other amino acids might be of less importance in the activation process of the ALK kinase than previously proposed. Thirdly, of the three individual tyrosines in the 1278-YRASYY-1283 activation loop, we find that Y1283 is the critical tyrosine in the activation process. Taken together, our observations employing different model systems reveal new mechanistic insights on how the full length ALK receptor is activated and highlight differences with earlier described activation mechanisms observed in the NPM-ALK fusion protein, supporting a mechanism of activation more in line with those observed for the Insulin Receptor (InR).

Place, publisher, year, edition, pages
MDPI , 2017. Vol. 9, no 11, article id 149
Keyword [en]
ALK, kinase activation, activation loop, gain-of-function, mutation analyses, Insulin receptor, NPM-ALK, oncogene, neuroblastoma
National Category
Cell and Molecular Biology
Identifiers
URN: urn:nbn:se:umu:diva-106700DOI: 10.3390/cancers9110149ISI: 000416603300005OAI: oai:DiVA.org:umu-106700DiVA, id: diva2:845379
Note

Originally included in thesis in manuscript form.

Available from: 2015-08-11 Created: 2015-08-03 Last updated: 2018-01-11Bibliographically approved
In thesis
1. Mechanistic Implications and Characterization of Anaplastic Lymphoma Kinase (ALK) mutations in Neuroblastoma
Open this publication in new window or tab >>Mechanistic Implications and Characterization of Anaplastic Lymphoma Kinase (ALK) mutations in Neuroblastoma
2015 (English)Doctoral thesis, comprehensive summary (Other academic)
Abstract [en]

Anaplastic lymphoma kinase (ALK) is a receptor tyrosine kinase that was first reported as a fusion partner of nucleophosmin in Anaplastic large cell lymphoma in 1994. ALK is involved in myriad of cancers including neuroblastoma which is the most common extracranial solid tumor affecting young children. It arises in the neural crest cells of sympathetic nervous system origin and is responsible for 12% of all childhood cancer deaths. Several point mutations in ALK have been described in both familial and sporadic neuroblastoma.

With the aim to understand the role of ALK in neuroblastoma further, we investigated the point mutations in ALK reported in patients. Using cell culture based methods and Drosophila as a model organism; we first characterized these mutations under three broad categories: 1) Ligand independent mutations that were constitutively active, 2) Kinase dead mutation and 3) Ligand dependent mutations that behaved as inducible wild type. Further, to understand the activation mechanism of ALK, we constructed mutations that could potentially alter ALK’s conformation based on the available crystal structure. From the data generated, we were able to provide a new perspective to the activation of full length ALK receptor. This was more in line with activation mechanism of insulin receptor and different from that suggested for ALK fusion protein. From a clinical point of view, all the mutations in the study were blocked to different degrees using the ALK inhibitor, crizotinib. Lastly, we identified potential downstream targets of ALK using phosphoproteomics. From the various targets identified, we focused on STAT3 and confirmed its role as a mediator in ALK initiated MYCN transcription. We showed that STAT3 inhibition led to reduction of MYCN levels and thereby identifying it as a potential therapeutic target in neuroblastoma. Overall, our study highlights clinical relevance of ALK mutations in neuroblastoma and from a basic biology viewpoint; it reveals important mechanistic insight into receptor activation.

Place, publisher, year, edition, pages
Umeå: Umeå Universitet, 2015. p. 79
Series
Umeå University medical dissertations, ISSN 0346-6612 ; 1709
Keyword
neuroblastoma, ALK, crizotinib, receptor tyrosine kinase
National Category
Cell and Molecular Biology
Research subject
Molecular Biology
Identifiers
urn:nbn:se:umu:diva-106663 (URN)978-91-7601-254-3 (ISBN)
Public defence
2015-10-02, KBC-huset, Stora hörsalen, KKB3B1, Umeå universitet, Umeå, 09:00 (English)
Opponent
Supervisors
Available from: 2015-09-04 Created: 2015-07-28 Last updated: 2018-01-11Bibliographically approved

Open Access in DiVA

fulltext(4275 kB)12 downloads
File information
File name FULLTEXT01.pdfFile size 4275 kBChecksum SHA-512
583b48f6702274facd7828909c3852928b0e08562cfb2f969da11d293ceb37463a8e1ef2642ff3c59709850034ce6345d3638b8e377769e0e8149a51e3649282
Type fulltextMimetype application/pdf

Other links

Publisher's full text

Search in DiVA

By author/editor
Chand, DaminiRuuth, KristinaHallberg, Bengt
By organisation
Department of Molecular Biology (Faculty of Medicine)
In the same journal
Cancers
Cell and Molecular Biology

Search outside of DiVA

GoogleGoogle Scholar
Total: 12 downloads
The number of downloads is the sum of all downloads of full texts. It may include eg previous versions that are now no longer available

doi
urn-nbn

Altmetric score

doi
urn-nbn
Total: 304 hits
CiteExportLink to record
Permanent link

Direct link
Cite
Citation style
  • apa
  • ieee
  • modern-language-association-8th-edition
  • vancouver
  • Other style
More styles
Language
  • de-DE
  • en-GB
  • en-US
  • fi-FI
  • nn-NO
  • nn-NB
  • sv-SE
  • Other locale
More languages
Output format
  • html
  • text
  • asciidoc
  • rtf