Molecular Cloning and Sequencing of a 58-kDa Membrane- and Microfilament-associated Protein from Ascites Tumor Cell Microvilli with Sequence Similarities to Retroviral Gag Proteins

Shin Hun Juang, Jiaqui Huang, Yongqing Li, Pedro J Salas, Nevis L. Fregien, Coralie A. Carothers Carraway, Kermit L. Carraway

Research output: Contribution to journalArticle

13 Citations (Scopus)

Abstract

The MAT-C1 subline of the 13762 rat mammary adenocarcinoma has highly stable, branched microvilli and immobile cell surface receptors. A membrane- and microfilament-associated 58-kDa protein (p58) in the MAT-C1 microvilli has been implicated in the stabilization of the microvilli and microfilament-membrane interactions. This protein is associated with a high Mr glycoprotein complex containing the (proto)oncogene p185neu and other signal transduction components in a putative microfilament-associated signal transduction particle. Amino acid sequences were obtained from two trypsin peptides of p58. Screening a MAT-C1 cDNA library with a degenerate oligonucleotide derived from the larger peptide and polymerase chain reaction amplification of cDNA ends permitted the isolation of overlapping cDNAs encoding the 427-amino acid open reading frame of p58. In vitro transcription and translation using a full-length cDNA gave a protein of approximately 55 kDa, which reacts with anti-p58 antiserum and reconstitutes into a complex with actin and glycoproteins from the membrane-microfilament interaction site. When COS-7 cells were transfected with the full-length cDNA, p58 was localized in a punctate distribution. In addition, the transfected cells exhibited fewer microfilament cables than untransfected neighboring cells. The amino acid sequence showed a surprising similarity to mammalian retroviral Gag proteins and included regions corresponding to p15, p12 and the N-terminal 80% of p30. Comparisons of p58 and the corresponding regions of the Gag proteins for Moloney murine leukemia virus indicated that about 60% of their amino acid residues were identical. These studies suggest that p58 is the product of an endogenous retroviral gene whose expression as a cellular protein alters the properties of the tumor cell to provide a selective advantage for tumor growth in the animal.

Original languageEnglish
Pages (from-to)15067-15075
Number of pages9
JournalJournal of Biological Chemistry
Volume269
Issue number21
StatePublished - May 27 1994

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gag Gene Products
Microfilament Proteins
Cloning
Molecular Cloning
Microvilli
Actin Cytoskeleton
Ascites
Tumors
Membrane Proteins
Complementary DNA
Cells
Membranes
Amino Acids
Signal transduction
Neoplasms
Proteins
Amino Acid Sequence
Signal Transduction
Peptides
Polymerase chain reaction

ASJC Scopus subject areas

  • Biochemistry

Cite this

Molecular Cloning and Sequencing of a 58-kDa Membrane- and Microfilament-associated Protein from Ascites Tumor Cell Microvilli with Sequence Similarities to Retroviral Gag Proteins. / Juang, Shin Hun; Huang, Jiaqui; Li, Yongqing; Salas, Pedro J; Fregien, Nevis L.; Carothers Carraway, Coralie A.; Carraway, Kermit L.

In: Journal of Biological Chemistry, Vol. 269, No. 21, 27.05.1994, p. 15067-15075.

Research output: Contribution to journalArticle

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title = "Molecular Cloning and Sequencing of a 58-kDa Membrane- and Microfilament-associated Protein from Ascites Tumor Cell Microvilli with Sequence Similarities to Retroviral Gag Proteins",
abstract = "The MAT-C1 subline of the 13762 rat mammary adenocarcinoma has highly stable, branched microvilli and immobile cell surface receptors. A membrane- and microfilament-associated 58-kDa protein (p58) in the MAT-C1 microvilli has been implicated in the stabilization of the microvilli and microfilament-membrane interactions. This protein is associated with a high Mr glycoprotein complex containing the (proto)oncogene p185neu and other signal transduction components in a putative microfilament-associated signal transduction particle. Amino acid sequences were obtained from two trypsin peptides of p58. Screening a MAT-C1 cDNA library with a degenerate oligonucleotide derived from the larger peptide and polymerase chain reaction amplification of cDNA ends permitted the isolation of overlapping cDNAs encoding the 427-amino acid open reading frame of p58. In vitro transcription and translation using a full-length cDNA gave a protein of approximately 55 kDa, which reacts with anti-p58 antiserum and reconstitutes into a complex with actin and glycoproteins from the membrane-microfilament interaction site. When COS-7 cells were transfected with the full-length cDNA, p58 was localized in a punctate distribution. In addition, the transfected cells exhibited fewer microfilament cables than untransfected neighboring cells. The amino acid sequence showed a surprising similarity to mammalian retroviral Gag proteins and included regions corresponding to p15, p12 and the N-terminal 80{\%} of p30. Comparisons of p58 and the corresponding regions of the Gag proteins for Moloney murine leukemia virus indicated that about 60{\%} of their amino acid residues were identical. These studies suggest that p58 is the product of an endogenous retroviral gene whose expression as a cellular protein alters the properties of the tumor cell to provide a selective advantage for tumor growth in the animal.",
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T1 - Molecular Cloning and Sequencing of a 58-kDa Membrane- and Microfilament-associated Protein from Ascites Tumor Cell Microvilli with Sequence Similarities to Retroviral Gag Proteins

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AU - Li, Yongqing

AU - Salas, Pedro J

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