Activity profiling of deubiquitinating enzymes in cervical carcinoma biopsies and cell lines.
Ubiquitin specific proteases (USPs) regulate the production and recycling of ubiquitin and are thereby critically involved in the control of cell growth, differentiation, and apoptosis. Increasing evidence implicates deregulation of USPs in malignant transformation but there is very little information on the overall and specific activity of USPs in normal and tumor tissues. We have used a chemistry-based functional proteomics approach to profile the activities of individual USPs in biopsies of human papillomavirus (HPV) carrying cervical carcinoma and adjacent normal tissue. To assess the contribution of HPV proteins, USP activity was also compared in HPV positive and negative cervical carcinoma cell lines and HPV E6/E7 immortalized human keratinocytes. The activity of the C-terminal hydrolases UCH-L3 and UCH37 was upregulated in the majority of tumor tissues compared to the adjacent normal tissues. UCH-L1 activity was lower in a significant proportion of the tumors but to a less extent in advanced tumors. In accordance with the relatively low UCH-L1 activity in tumor biopsies, UCH-L1 was detected only in one out of eight cervical carcinoma lines. UCH-L1, UCH-L3, USP7, and USP9X activity was upregulated following HPV E6/E7 immortalization of keratinocytes, suggesting a role of these enzymes in growth transformation.
Rolén
Ulrika
U
Microbiology and Tumor Biology Center, Karolinska Institutet, Stockholm, Sweden.
Kobzeva
Vera
V
Gasparjan
Natalja
N
Ovaa
Huib
H
Winberg
Gösta
G
Kisseljov
Fjodor
F
Masucci
Maria G
MG
D001706:Biopsy; D002268:Carboxypeptidases; D002294:Carcinoma, Squamous Cell / Q000201:enzymology / Q000473:pathology / Q000821:virology; D002352:Carrier Proteins / Q000378:metabolism*; D016022:Case-Control Studies; D002460:Cell Line; D002472:Cell Transformation, Viral / Q000235:genetics; D002584:Cervix Uteri / Q000201:enzymology / Q000473:pathology / Q000821:virology; D004279:DNA, Viral / Q000235:genetics; D004268:DNA-Binding Proteins / Q000235:genetics; D005260:Female; D006801:Humans; D015603:Keratinocytes / Q000166:cytology / Q000201:enzymology / Q000821:virology; D008207:Lymphatic Metastasis / Q000473:pathology; D009856:Oncogene Proteins, Viral / Q000235:genetics; D027383:Papillomaviridae / Q000302:isolation & purification; D030361:Papillomavirus Infections / Q000201:enzymology / Q000473:pathology / Q000821:virology; D012097:Repressor Proteins / Q000235:genetics; D043222:Ubiquitin Thiolesterase / Q000378:metabolism*; D002583:Uterine Cervical Neoplasms / Q000201:enzymology* / Q000473:pathology / Q000821:virology
D002352:Carrier Proteins; D004279:DNA, Viral; D004268:DNA-Binding Proteins; C068651:E6 protein, Human papillomavirus type 16; C052603:E6 protein, Human papillomavirus type 18; C052633:E7 protein, Human papillomavirus type 18; D009856:Oncogene Proteins, Viral; D012097:Repressor Proteins; C482860:UCHL1 protein, human; D002268:Carboxypeptidases; C421245:UCHL5 protein, human; D043222:Ubiquitin Thiolesterase