Journal Article

The small tumor antigen of Merkel cell polyomavirus accomplishes cellular transformation by uniquely localizing to the nucleus despite the absence of a known nuclear localization signal

Thevenin, Kaira R., Schrager, Matt, Di Benedetto, Cody E., Tieche, Isabella S., Dye, Kristine N.

2024

Overview

Background Merkel Cell Carcinoma (MCC) is an aggressive skin cancer that is three times deadlier than melanoma. In 2008, it was found that 80% of MCC cases are caused by the genomic integration of a novel polyomavirus, Merkel Cell Polyomavirus (MCPyV), and the expression of its small and truncated large tumor antigens (ST and LT-t, respectively). MCPyV belongs to a family of human polyomaviruses; however, it is the only one with a clear association to cancer. Methods To investigate the role and mechanisms of various polyomavirus tumor antigens in cellular transformation, Rat-2 and 293A cells were transduced with pLENTI MCPyV LT-t, MCPyV ST, TSPyV ST, HPyV7 ST, or empty pLENTI and assessed through multiple transformation assays, and subcellular fractionations. One-way ANOVA tests were used to assess statistical significance. Results Soft agar, proliferation, doubling time, glucose uptake, and serum dependence assays confirmed ST to be the dominant transforming protein of MCPyV. Furthermore, it was found that MCPyV ST is uniquely transforming, as the ST antigens of other non-oncogenic human polyomaviruses such as Trichodysplasia Spinulosa-Associated Polyomavirus (TSPyV) and Human Polyomavirus 7 (HPyV7) were not transforming when similarly assessed. Identification of structural dissimilarities between transforming and non-transforming tumor antigens revealed that the uniquely transforming domain(s) of MCPyV ST are likely located within the structurally dissimilar loops of the MCPyV ST unique region. Of all known MCPyV ST cellular interactors, 62% are exclusively or transiently nuclear, suggesting that MCPyV ST localizes to the nucleus despite the absence of a canonical nuclear localization signal. Indeed, subcellular fractionations confirmed that MCPyV ST could achieve nuclear localization through a currently unknown, regulated mechanism independent of its small size, as HPyV7 and TSPyV ST proteins were incapable of nuclear translocation. Although nuclear localization was found to be important for several transforming properties of MCPyV ST, some properties were also performed by a cytoplasmic sequestered MCPyV ST, suggesting that MCPyV ST may perform different transforming functions in individual subcellular compartments. Conclusions Together, these data further elucidate the unique differences between MCPyV ST and other polyomavirus ST proteins necessary to understand MCPyV as the only known human oncogenic polyomavirus.

• Publisher: BioMed Central,BioMed Central Ltd,BMC
• Subject: agar / Animals / Antigens, Polyomavirus Transforming - genetics / Antigens, Polyomavirus Transforming - metabolism / Antigens, Viral, Tumor - genetics / Antigens, Viral, Tumor - metabolism / Biomedical and Life Sciences / Biomedicine / Biotechnology industry / blood serum / carcinoma / Carcinoma, Merkel Cell - virology / Care and treatment / Cell Line / Cell Nucleus - metabolism / Cell Nucleus - virology / Cell Transformation, Viral / Cellular transformation / Dextrose / Diagnosis / domain / family / genomics / Glucose / Human polyomavirus 5 / Human polyomavirus 7 / Human polyomavirus 8 / Humans / Immunosuppression / Melanoma / Merkel cell carcinoma / Merkel cell polyomavirus / Merkel cell polyomavirus - genetics / Merkel cell polyomavirus - physiology / neoplasm antigens / Nuclear localization / Nuclear Localization Signals / Patient outcomes / Polyomavirus Infections - virology / Proteins / Rats / Risk factors / Skin / Skin cancer / skin neoplasms / Skin Neoplasms - pathology / Skin Neoplasms - virology / Testing / Trichodysplasia spinulosa polyomavirus / Tumor antigens / Virology