title: Molecular cloning of NILE glycoprotein and evidence for its continued expression in mature rat CNS.
abstract: The NILE glycoprotein is a rat neuronal cell adhesion molecule which has been reported to be very similar in structure, function, and distribution to the mouse L1 glycoprotein. Here we report the complete nucleotide sequence of the NILE message (5,208 nucleotides) and the deduced amino acid sequence of the NILE polypeptide (1,257 amino acids). The predicted NILE protein is 96% identical to L1 at the amino acid level, confirming that the two molecules are homologues. The sequence information shows that NILE is a transmembrane molecule with an extensive ectodomain and a much smaller cytoplasmic domain. The extracellular portion of the molecule contains six immunoglobulin C-2 type domains followed by five fibronectin type III repeats. These two structural motifs are characteristic of several other cell adhesion molecules. The cytoplasmic tails of NILE and L1 are identical to each other and distinct from the cytoplasmic regions of all other cell adhesion molecules except Ng-CAM and neuroglian. Several possible sites for phosphorylation are present in the cytoplasmic tail of NILE. Antisera were produced against two NILE-beta-galactosidase fusion proteins containing distinct segments of the NILE polypeptide: the cytoplasmic domain and the segment containing fibronectin type III repeats. Immunoblots with these antisera and Northern blots with a NILE cDNA probe indicate that NILE continues to be expressed in most areas of the mature rat brain. This contradicts previous immunofluorescence data, which suggested that NILE was substantially down-regulated in maturing nerve fiber tracts. This raises the possibility that NILE could be masked in situ by interactions with other cell surface molecules.
authors.affiliation: La Jolla Cancer Research Foundation, California 92037.
authors.forename: J T
authors.initials: JT
authors.lastname: Prince
authors.forename: L
authors.initials: L
authors.lastname: Alberti
authors.forename: P A
authors.initials: PA
authors.lastname: Healy
authors.forename: S J
authors.initials: SJ
authors.lastname: Nauman
authors.forename: W B
authors.initials: WB
authors.lastname: Stallcup
mesh_terms: D000375:Aging; D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D001921:Brain / Q000254:growth & development / Q000502:physiology*; D003001:Cloning, Molecular / Q000379:methods; D004273:DNA, Neoplasm / Q000235:genetics / Q000302:isolation & purification; D015870:Gene Expression; D015723:Gene Library; D008562:Membrane Glycoproteins / Q000096:biosynthesis / Q000235:genetics* / Q000302:isolation & purification; D008969:Molecular Sequence Data; D009414:Nerve Growth Factors / Q000235:genetics; D039842:Neural Cell Adhesion Molecule L1; D009928:Organ Specificity; D016716:PC12 Cells; D016133:Polymerase Chain Reaction / Q000379:methods; D012333:RNA, Messenger / Q000235:genetics; D051381:Rats; D011919:Rats, Inbred Strains; D011993:Recombinant Fusion Proteins / Q000096:biosynthesis / Q000302:isolation & purification; D013116:Spinal Cord / Q000254:growth & development / Q000502:physiology*; D014158:Transcription, Genetic; D001616:beta-Galactosidase / Q000096:biosynthesis / Q000235:genetics / Q000302:isolation & purification
chemical_list: D004273:DNA, Neoplasm; D008562:Membrane Glycoproteins; D009414:Nerve Growth Factors; D039842:Neural Cell Adhesion Molecule L1; D012333:RNA, Messenger; D011993:Recombinant Fusion Proteins; D001616:beta-Galactosidase