Nucleolin (NCL) is a major component of the cell nucleolus, which has the ability to rapidly shuttle to several other cell’s compartments. NCL has been shown to play important roles in a variety of essential functions among which ribosome biogenesis, gene expression and cell growth. However, the precise mechanisms underlying NCL functions are still unclear. Our study aimed to provide new information on NCL functions via the identification of its nuclear interacting partners. Using an interactomics approach, we identified 140 binding partners of NCL, among which, a 100 of them, were specifically found associated to NCL independently of the presence of RNA.