Limits...
Differential reprogramming of isogenic colorectal cancer cells by distinct activating KRAS mutations.

Hammond DE, Mageean CJ, Rusilowicz EV, Wickenden JA, Clague MJ, Prior IA - J. Proteome Res. (2015)

Bottom Line: Oncogenic mutations of Ras at codons 12, 13, or 61, that render the protein constitutively active, are found in ∼ 16% of all cancer cases.To investigate the differential effects upon cell status associated with KRAS mutations we performed a quantitative analysis of the proteome and phosphoproteome of isogenic SW48 colon cancer cell lines in which one allele of the endogenous gene has been edited to harbor specific KRAS mutations (G12V, G12D, or G13D).One notable example of specific up-regulation in KRAS codon 12 mutant SW48 cells is provided by the short form of the colon cancer stem cell marker doublecortin-like Kinase 1 (DCLK1) that can be reversed by suppression of KRAS.

View Article: PubMed Central - PubMed

Affiliation: Division of Cellular and Molecular Physiology, Institute of Translational Medicine, University of Liverpool , Crown Street, Liverpool L69 3BX, United Kingdom.

ABSTRACT
Oncogenic mutations of Ras at codons 12, 13, or 61, that render the protein constitutively active, are found in ∼ 16% of all cancer cases. Among the three major Ras isoforms, KRAS is the most frequently mutated isoform in cancer. Each Ras isoform and tumor type displays a distinct pattern of codon-specific mutations. In colon cancer, KRAS is typically mutated at codon 12, but a significant fraction of patients have mutations at codon 13. Clinical data suggest different outcomes and responsiveness to treatment between these two groups. To investigate the differential effects upon cell status associated with KRAS mutations we performed a quantitative analysis of the proteome and phosphoproteome of isogenic SW48 colon cancer cell lines in which one allele of the endogenous gene has been edited to harbor specific KRAS mutations (G12V, G12D, or G13D). Each mutation generates a distinct signature, with the most variability seen between G13D and the codon 12 KRAS mutants. One notable example of specific up-regulation in KRAS codon 12 mutant SW48 cells is provided by the short form of the colon cancer stem cell marker doublecortin-like Kinase 1 (DCLK1) that can be reversed by suppression of KRAS.

No MeSH data available.


Related in: MedlinePlus

Relationship between individual phosphopeptide responsesand proteomechanges. The top 15 proteins with multiple phosphosites measured inthe combined MaxQuant analysis and a minimum of one phosphopeptidewithin cluster 5 or 6 are collated together with their respectiveproteome values. Ratios for both codon 12 mutations versus G13D aredisplayed as indicated. Darker red or blue spots indicate multipleoverlapping responses.
© Copyright Policy
Related In: Results  -  Collection

License
getmorefigures.php?uid=PMC4356034&req=5

fig4: Relationship between individual phosphopeptide responsesand proteomechanges. The top 15 proteins with multiple phosphosites measured inthe combined MaxQuant analysis and a minimum of one phosphopeptidewithin cluster 5 or 6 are collated together with their respectiveproteome values. Ratios for both codon 12 mutations versus G13D aredisplayed as indicated. Darker red or blue spots indicate multipleoverlapping responses.

Mentions: Phosphopeptide members of clusters 5 and 6 that originatedfromproteins with multiple phosphorylation sites were curated to examinethe patterns of response across all detected sites within these proteins(Figure 4 and SupportingInformation Figure 3). Where available, proteome data are alsopresented (squares) to see the extent to which phosphopeptide responseswere influenced by changes in protein abundance rather than a proportionalincrease in phosphorylation. In the majority of cases where comparisonscould be made, proteome changes were a minor influence on phosphopeptideratios. Interestingly, most phosphosites within a protein trendedin a similar direction for both cluster 5 and cluster 6 members, indicatingcoordinated increase or decrease of phosphorylation at multiple siteswithin a protein.


Differential reprogramming of isogenic colorectal cancer cells by distinct activating KRAS mutations.

Hammond DE, Mageean CJ, Rusilowicz EV, Wickenden JA, Clague MJ, Prior IA - J. Proteome Res. (2015)

Relationship between individual phosphopeptide responsesand proteomechanges. The top 15 proteins with multiple phosphosites measured inthe combined MaxQuant analysis and a minimum of one phosphopeptidewithin cluster 5 or 6 are collated together with their respectiveproteome values. Ratios for both codon 12 mutations versus G13D aredisplayed as indicated. Darker red or blue spots indicate multipleoverlapping responses.
© Copyright Policy
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4356034&req=5

fig4: Relationship between individual phosphopeptide responsesand proteomechanges. The top 15 proteins with multiple phosphosites measured inthe combined MaxQuant analysis and a minimum of one phosphopeptidewithin cluster 5 or 6 are collated together with their respectiveproteome values. Ratios for both codon 12 mutations versus G13D aredisplayed as indicated. Darker red or blue spots indicate multipleoverlapping responses.
Mentions: Phosphopeptide members of clusters 5 and 6 that originatedfromproteins with multiple phosphorylation sites were curated to examinethe patterns of response across all detected sites within these proteins(Figure 4 and SupportingInformation Figure 3). Where available, proteome data are alsopresented (squares) to see the extent to which phosphopeptide responseswere influenced by changes in protein abundance rather than a proportionalincrease in phosphorylation. In the majority of cases where comparisonscould be made, proteome changes were a minor influence on phosphopeptideratios. Interestingly, most phosphosites within a protein trendedin a similar direction for both cluster 5 and cluster 6 members, indicatingcoordinated increase or decrease of phosphorylation at multiple siteswithin a protein.

Bottom Line: Oncogenic mutations of Ras at codons 12, 13, or 61, that render the protein constitutively active, are found in ∼ 16% of all cancer cases.To investigate the differential effects upon cell status associated with KRAS mutations we performed a quantitative analysis of the proteome and phosphoproteome of isogenic SW48 colon cancer cell lines in which one allele of the endogenous gene has been edited to harbor specific KRAS mutations (G12V, G12D, or G13D).One notable example of specific up-regulation in KRAS codon 12 mutant SW48 cells is provided by the short form of the colon cancer stem cell marker doublecortin-like Kinase 1 (DCLK1) that can be reversed by suppression of KRAS.

View Article: PubMed Central - PubMed

Affiliation: Division of Cellular and Molecular Physiology, Institute of Translational Medicine, University of Liverpool , Crown Street, Liverpool L69 3BX, United Kingdom.

ABSTRACT
Oncogenic mutations of Ras at codons 12, 13, or 61, that render the protein constitutively active, are found in ∼ 16% of all cancer cases. Among the three major Ras isoforms, KRAS is the most frequently mutated isoform in cancer. Each Ras isoform and tumor type displays a distinct pattern of codon-specific mutations. In colon cancer, KRAS is typically mutated at codon 12, but a significant fraction of patients have mutations at codon 13. Clinical data suggest different outcomes and responsiveness to treatment between these two groups. To investigate the differential effects upon cell status associated with KRAS mutations we performed a quantitative analysis of the proteome and phosphoproteome of isogenic SW48 colon cancer cell lines in which one allele of the endogenous gene has been edited to harbor specific KRAS mutations (G12V, G12D, or G13D). Each mutation generates a distinct signature, with the most variability seen between G13D and the codon 12 KRAS mutants. One notable example of specific up-regulation in KRAS codon 12 mutant SW48 cells is provided by the short form of the colon cancer stem cell marker doublecortin-like Kinase 1 (DCLK1) that can be reversed by suppression of KRAS.

No MeSH data available.


Related in: MedlinePlus