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Characterization and identification of PARM-1 as a new potential oncogene.

Charfi C, Levros LC, Edouard E, Rassart E - Mol. Cancer (2013)

Bottom Line: Moreover, deletion mutants of human PARM-1 without either extracellular or cytoplasmic portions seem to retain the ability to induce anchorage-independent growth of NIH/3T3 cells.In addition, PARM-1 increases ERK1/2, but more importantly AKT and STAT3 phosphorylation.Our results strongly suggest the oncogenic potential of PARM-1.

View Article: PubMed Central - HTML - PubMed

Affiliation: Laboratoire de Biologie Moléculaire, Département des Sciences Biologiques, Centre BioMed, Université du Québec à Montréal, Case Postale 8888, Succursale Centre-ville, Montréal, QC H3C-3P8, Canada.

ABSTRACT

Background: The Graffi murine retrovirus is a powerful tool to find leukemia associated oncogenes. Using DNA microarrays, we recently identified several genes specifically deregulated in T- and B-leukemias induced by this virus.

Results: In the present study, probsets associated with T-CD8+ leukemias were analyzed and we validated the expression profile of the Parm-1 gene. PARM-1 is a member of the mucin family. We showed that human PARM-1 is an intact secreted protein accumulating predominantly, such as murine PARM-1, at the Golgi and in the early and late endosomes. PARM-1 colocalization with α-tubulin suggests that its trafficking within the cell involves the microtubule cytoskeleton. Also, the protein co-localizes with caveolin-1 which probably mediates its internalization. Transient transfection of both mouse and human Parm-1 cDNAs conferred anchorage- and serum-independent growth and enhanced cell proliferation. Moreover, deletion mutants of human PARM-1 without either extracellular or cytoplasmic portions seem to retain the ability to induce anchorage-independent growth of NIH/3T3 cells. In addition, PARM-1 increases ERK1/2, but more importantly AKT and STAT3 phosphorylation.

Conclusions: Our results strongly suggest the oncogenic potential of PARM-1.

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Related in: MedlinePlus

Schematic representation of full-length and deletion mutant constructs of mPARM-1 and hPARM-1. (a and b) Full-length constructs of the mouse (296 a.a) and human PARM-1 (310 a.a). (c to g) representation of the human deletion mutants. SP: signal peptide: (1–20 a.a), EC: extracellular domain (mPARM-1: 20–246 a.a; hPARM-1: 20–260 a.a), TM: transmembrane domain (mPARM-1: 246–266 a.a; hPARM-1: 260–280 a.a), and CT: cytoplasmic tail (mPARM-1: 266–296 a.a; hPARM-1: 280–310 a.a). EGFP was fused in the C-terminal end of all constructs.
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Figure 2: Schematic representation of full-length and deletion mutant constructs of mPARM-1 and hPARM-1. (a and b) Full-length constructs of the mouse (296 a.a) and human PARM-1 (310 a.a). (c to g) representation of the human deletion mutants. SP: signal peptide: (1–20 a.a), EC: extracellular domain (mPARM-1: 20–246 a.a; hPARM-1: 20–260 a.a), TM: transmembrane domain (mPARM-1: 246–266 a.a; hPARM-1: 260–280 a.a), and CT: cytoplasmic tail (mPARM-1: 266–296 a.a; hPARM-1: 280–310 a.a). EGFP was fused in the C-terminal end of all constructs.

Mentions: PARM-1 is a member of the mucin family known to be expressed at the surface of many epithelial cells [13] to promote cell survival by protecting the cell surface and to be implicated in cancer development [14]. Protein sequence analysis of mPARM-1 showed that, as the hPARM-1 and in addition to its single transmembrane domain, mPARM-1 possess an N-terminal signal peptide (Figure 2a and 2b) [15]. mPARM-1 sequence contains 3 N-glycosylated motifs and 65 mucin-type O-glycosylated sites [16], suggesting that, as its human counterpart, mPARM-1 should be highly glycosylated. Moreover, we found that 41% of the amino acid composition of mPARM-1 is represented by serine, proline and threonine residues similar to the human protein [17]. Interestingly, amino acid sequence alignment of PARM-1 homologs showed that the C-terminus is highly conserved (Additional file 1: Figure S1) suggesting an important role through evolution.


Characterization and identification of PARM-1 as a new potential oncogene.

Charfi C, Levros LC, Edouard E, Rassart E - Mol. Cancer (2013)

Schematic representation of full-length and deletion mutant constructs of mPARM-1 and hPARM-1. (a and b) Full-length constructs of the mouse (296 a.a) and human PARM-1 (310 a.a). (c to g) representation of the human deletion mutants. SP: signal peptide: (1–20 a.a), EC: extracellular domain (mPARM-1: 20–246 a.a; hPARM-1: 20–260 a.a), TM: transmembrane domain (mPARM-1: 246–266 a.a; hPARM-1: 260–280 a.a), and CT: cytoplasmic tail (mPARM-1: 266–296 a.a; hPARM-1: 280–310 a.a). EGFP was fused in the C-terminal end of all constructs.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC3750824&req=5

Figure 2: Schematic representation of full-length and deletion mutant constructs of mPARM-1 and hPARM-1. (a and b) Full-length constructs of the mouse (296 a.a) and human PARM-1 (310 a.a). (c to g) representation of the human deletion mutants. SP: signal peptide: (1–20 a.a), EC: extracellular domain (mPARM-1: 20–246 a.a; hPARM-1: 20–260 a.a), TM: transmembrane domain (mPARM-1: 246–266 a.a; hPARM-1: 260–280 a.a), and CT: cytoplasmic tail (mPARM-1: 266–296 a.a; hPARM-1: 280–310 a.a). EGFP was fused in the C-terminal end of all constructs.
Mentions: PARM-1 is a member of the mucin family known to be expressed at the surface of many epithelial cells [13] to promote cell survival by protecting the cell surface and to be implicated in cancer development [14]. Protein sequence analysis of mPARM-1 showed that, as the hPARM-1 and in addition to its single transmembrane domain, mPARM-1 possess an N-terminal signal peptide (Figure 2a and 2b) [15]. mPARM-1 sequence contains 3 N-glycosylated motifs and 65 mucin-type O-glycosylated sites [16], suggesting that, as its human counterpart, mPARM-1 should be highly glycosylated. Moreover, we found that 41% of the amino acid composition of mPARM-1 is represented by serine, proline and threonine residues similar to the human protein [17]. Interestingly, amino acid sequence alignment of PARM-1 homologs showed that the C-terminus is highly conserved (Additional file 1: Figure S1) suggesting an important role through evolution.

Bottom Line: Moreover, deletion mutants of human PARM-1 without either extracellular or cytoplasmic portions seem to retain the ability to induce anchorage-independent growth of NIH/3T3 cells.In addition, PARM-1 increases ERK1/2, but more importantly AKT and STAT3 phosphorylation.Our results strongly suggest the oncogenic potential of PARM-1.

View Article: PubMed Central - HTML - PubMed

Affiliation: Laboratoire de Biologie Moléculaire, Département des Sciences Biologiques, Centre BioMed, Université du Québec à Montréal, Case Postale 8888, Succursale Centre-ville, Montréal, QC H3C-3P8, Canada.

ABSTRACT

Background: The Graffi murine retrovirus is a powerful tool to find leukemia associated oncogenes. Using DNA microarrays, we recently identified several genes specifically deregulated in T- and B-leukemias induced by this virus.

Results: In the present study, probsets associated with T-CD8+ leukemias were analyzed and we validated the expression profile of the Parm-1 gene. PARM-1 is a member of the mucin family. We showed that human PARM-1 is an intact secreted protein accumulating predominantly, such as murine PARM-1, at the Golgi and in the early and late endosomes. PARM-1 colocalization with α-tubulin suggests that its trafficking within the cell involves the microtubule cytoskeleton. Also, the protein co-localizes with caveolin-1 which probably mediates its internalization. Transient transfection of both mouse and human Parm-1 cDNAs conferred anchorage- and serum-independent growth and enhanced cell proliferation. Moreover, deletion mutants of human PARM-1 without either extracellular or cytoplasmic portions seem to retain the ability to induce anchorage-independent growth of NIH/3T3 cells. In addition, PARM-1 increases ERK1/2, but more importantly AKT and STAT3 phosphorylation.

Conclusions: Our results strongly suggest the oncogenic potential of PARM-1.

Show MeSH
Related in: MedlinePlus