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(American Journal of Pathology. 2001;159:1363-1374.)
© 2001 American Society for Investigative Pathology

Regular Articles

Increased Expression of a Nucleolar Nop5/Sik Family Member in Metastatic Melanoma Cells

Evidence for Its Role in Nucleolar Sizing and Function

Ken’i Nakamoto^*, Akihiko Ito, Kenji Watabe, Yu-ichiro Koma^*, Hideo Asada, Kunihiko Yoshikawa, Yasuhisa Shinomura, Yuji Matsuzawa, Hiroshi Nojima^* and Yukihiko Kitamura

From the Department of Molecular Genetics,^*
Research Institute for Microbial Diseases, Osaka University, Suita, Osaka; and the Departments of Pathology,
Dermatology,
and Internal Medicine and Molecular Science,
Osaka University Medical School, Suita, Osaka, Japan

F10 and BL6 cells of B16 mouse melanoma cells are metastatic after intravenous injection, but only BL6 cells can metastasize to lungs after subcutaneous injection. Differences in gene expression between the two cell lines were examined, and a greater expression of the Sik-similar protein (Sik-SP) gene was found in BL6 cells. Structurally, Sik-SP belongs to the nucleolar Nop5/Sik family whose members play central roles in ribosome biogenesis; however, the function of Sik-SP has not been examined. Cytology with green fluorescent protein-fused proteins showed that Sik-SP was localized to the nucleolus. To examine whether Sik-SP is involved in ribosome biogenesis, two parameters were measured: magnitude of ribosomal RNA synthesis per nucleus and magnitude of protein production from the same amount of mRNA of an exogenous luciferase gene. Both values and, in addition, nucleolar size were larger in COS-7 monkey kidney cells overexpressing Sik-SP and BL6 cells than in mock-transfected COS-7 and F10 cells, respectively. Sik-SP seemed to promote ribosome biogenesis in the nucleolus. Furthermore, the expression of Sik-SP seemed to confer a greater cell growth response to serum, because such a response was greater in BL6 cells and F10 cells overexpressing Sik-SP than in untreated and mock-transfected F10 cells. Sik-SP may render melanoma cells more competent to survive through augmenting the activity of nucleolus.

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