Nucleolin is a protein that in humans is encoded by the NCL gene. Nucleolin is the major nucleolar protein of growing eukaryotic cells. It is found associated with intranucleolar chromatin and pre-ribosomal particles. It induces chromatin decondensation by binding to histone H1. It is thought to play a role in pre-rRNA transcription and ribosome assembly. May play a role in the process of transcriptional elongation. Binds RNA oligonucleotides with 5'-UUAGGG-3' repeats more tightly than the telomeric single-stranded DNA 5'-TTAGGG-3' repeats. Nucleolin is also able to act as a transcriptional coactivator with Chicken Ovalbumin Upstream Promoter Transcription Factor II (COUP-TFII).
Background References
1. "Structure and function of the N-terminal nucleolin binding domain of nuclear valosin-containing protein-like 2 (NVL2) harboring a nucleolar localization signal." Fujiwara Y., Fujiwara K., Goda N., Iwaya N., Tenno T., Shirakawa M., Hiroaki H. J. Biol. Chem. 286:21732-21741(2011) "Functional proteomic analysis of human nucleolus." Scherl A., Coute Y., Deon C., Calle A., Kindbeiter K., Sanchez J.-C., Greco A., Hochstrasser D.F., Diaz J.-J.Mol. Biol. Cell 13:4100-4109(2002)
Post-translational Modification
Some glutamate residues are glycylated by TTLL8. This modification occurs exclusively on glutamate residues and results in a glycine chain on the gamma-carboxyl group (By similarity).; Symmetrically methylated by PRMT5.
Western blot analysis of Nucleolin on different lysates with Rabbit anti-Nucleolin antibody (R1307-2) at 1/1,000 dilution.
Lane 1: HeLa cell lysate Lane 2: Jurkat cell lysate Lane 3: HEK-293 cell lysate Lane 4: A431 cell lysate Lane 5: HepG2 cell lysate Lane 6: A549 cell lysate
Lysates/proteins at 20 µg/Lane.
Predicted band size: 77 kDa Observed band size: 100 kDa
Exposure time: 1 minute;
4-20% SDS-PAGE gel.
Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody (R1307-2) at 1/1,000 dilution was used in 5% NFDM/TBST at room temperature for 2 hours. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1/100,000 dilution was used for 1 hour at room temperature.
ICC staining nucleolin in mouse embryonic stem cells (red). Cells were fixed in paraformaldehyde, permeabilised with 0.25% Triton X100/PBS and counterstained with DAPI in order to highlight the nucleus (blue).
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