Molecular cloning and sequencing of the mucin subunit of a heterodimeric, bifunctional cell surface glycoprotein complex of ascites rat mammary adenocarcinoma cells

• Published in: The Journal of biological chemistry 1994-04, Vol.269 (16), p.11950-11955
• Main Authors: WU, K, FREGIEN, N, CARRAWAY, K. L
• Format: Article
• Language: English
• Subjects: Adenocarcinoma - chemistry; Adenocarcinoma - metabolism; Amino Acid Sequence; Analytical, structural and metabolic biochemistry; Animals; Base Sequence; Biological and medical sciences; Cloning, Molecular; DNA Primers; Female; Fundamental and applied biological sciences. Psychology; Glycoproteins; Mammary Neoplasms, Experimental - metabolism; Membrane Glycoproteins - biosynthesis; Membrane Glycoproteins - chemistry; Molecular Sequence Data; Mucin-4; Neoplasm Proteins - biosynthesis; Polymerase Chain Reaction; Proteins; Rats; Repetitive Sequences, Nucleic Acid; RNA, Neoplasm - isolation & purification; RNA, Neoplasm - metabolism; Sequence Homology, Amino Acid; Sialoglycoproteins - biosynthesis; Sialoglycoproteins - chemistry
• ISSN: 0021-9258; 1083-351X
• DOI: 10.1016/S0021-9258(17)32665-0

Ascites sublines of the highly metastatic 13762 rat mammary adenocarcinoma contain abundant amounts of a heterodimeric cell surface glycoprotein complex composed of a mucin subunit ASGP-1 (ascites sialoglycoprotein-1) and a transmembrane subunit (ASGP-2). Previous studies showed that the complex is synthesized from a single polypeptide encoded by a 9 kb transcript. The sequence of the transmembrane subunit was obtained from a 5-kilobase (kb) cDNA isolated from a plasmid library (Sheng, Z., Wu, K., Carraway, K. L., and Fregien, N. (1992) J. Biol. Chem. 267, 16341-16346). Completion of the sequence of this cDNA revealed the C-terminal domain of ASGP-1, which is rich in serine and threonine but contains no typical mucin-type repeats. The remainder of the sequence of ASGP-1 and the 9-kb transcript was obtained by two 5'-RACE (rapid amplification of cDNA ends) steps and primer extension analysis. These results revealed that the 5' half of the 9-kb transcript contains a short 5'-noncoding region and encodes a signal sequence, a short nonrepeat region, and a repeat domain containing 11 repeats. Nine of these repeats are found in tandem, but the two end repeats are separated from the others by short unique sequences. The repeats vary from 117-124 amino acids and are 70-90% identical to a consensus sequence. Overall, the sequence predicts that ASGP-1 contains 2172 amino acids (M(r) 224,190), 43% of which are serine and threonine. We propose that the complex of this mucin and its transmembrane subunit, which contains growth factor-modulating activity, may play an important role in tumor progression.