NANOG/NANOGP8 Localizes at the Centrosome and is Spatiotemporally Associated with Centriole Maturation

Abstract

NANOG is a transcription factor involved in the regulation of pluripotency and stemness. The functional paralog of NANOG, NANOGP8, differs from NANOG in only three amino acids and exhibits similar reprogramming activity. Given the transcriptional regulatory role played by NANOG, the nuclear localization of NANOG/NANOGP8 has primarily been considered to date. In this study, we investigated the intriguing extranuclear localization of NANOG and demonstrated that a substantial pool of NANOG/NANOGP8 is localized at the centrosome. Using double immunofluorescence, the colocalization of NANOG protein with pericentrin was identified by two independent anti-NANOG antibodies among 11 tumor and non-tumor cell lines. The validity of these observations was confirmed by transient expression of GFP-tagged NANOG, which also colocalized with pericentrin. Mass spectrometry of the anti-NANOG immunoprecipitated samples verified the antibody specificity and revealed the expression of both NANOG and NANOGP8, which was further confirmed by real-time PCR. Using cell fractionation, we show that a considerable amount of NANOG protein is present in the cytoplasm of RD and NTERA-2 cells. Importantly, cytoplasmic NANOG was unevenly distributed at the centrosome pair during the cell cycle and colocalized with the distal region of the mother centriole, and its presence was markedly associated with centriole maturation. Along with the finding that the centrosomal localization of NANOG/NANOGP8 was detected in various tumor and non-tumor cell types, these results provide the first evidence suggesting a common centrosome-specific role of NANOG.

Nanog, Stat-3, and Sox-5 involvement in human fetal temporomandibular joint late development

“… 22 Recently, cytoplasmic, more precisely centrosome localization of Nanog, during the cell cycle was detected in various tumor and non-tumor cell types. 23 Fan et al, 2021 24 recently described a fibrocartilage stem cell population in a fibrous superficial zone of the adult human mandibular condyle. The formation of these cells occurs during the late period of the embryonic stage, 25 which corroborate with our data.…”

RNAi mediated silencing of Nanog expression suppresses the growth of human colorectal cancer stem cells

“…The functional paralog of Nanog, NanogP8, differs from Nanog in only three amino acids and exhibits similar reprogramming activity. E. Mikulenkova et al investigated the intriguing extranuclear localization of Nanog and demonstrated that a substantial pool of Nanog/NanogP8 is localized at the centrosome [31]. NanogP8 is the main regulator of gastric cancer stem cells.…”

The Centrosome: Conclusions and Perspectives

“…Similarly, Mikulenkova, together with the paper’s co-authors, found that the nuclear protein NANOG/NANOGP8 is localized in the centrosome [ 2 ]. The above examples show that various regulatory pathways intersect in the centrosome; thus, regulatory proteins are concentrated in the centrosome.…”