Characterization of human SHC p66 cdna and its processed pseudogene mapping to Xq12-q13.1

Roslan B. Harun, Kim K. Smith, Jack P. Leek, Alexander F. Markham, Alan Morris, John F J Morrison

Research output: Contribution to journalArticle

4 Citations (Scopus)

Abstract

SHC is an adapter protein in the Ras-MAPkinase pathway that is involved in the regulation of cell growth and differentiation. The p46 and p52 isoforms are thought to be produced by the use of two alternative translation initiation sites in a 3.4-kb transcript from the SHCA gene, which maps to chromosome 1q21. The p68 isoform could be encoded by a different 3.8-or 2.8- kb transcript of the same gene or alternatively by a SHC-related gone. To characterize other putative genes coding for SHC-like proteins, primers from the 3' UTR of the SHCA gene were used to screen a yeast artificial chromosome (YAC) library by polymerase chain reaction (PCR). Two YAC clones, 20D11B and 36D1D, were isolated and used as probes for fluorescence in situ hybridization analysis. Both these probes hybridized to chromosome Xq12- q13.1. This novel SHC-related sequence was characterized by direct sequencing of vectorette library PCR products produced from clone 20D11B. A transcript of 3.2 kb that was 85% identical to the mouse Shc cDNA encoding the p66 isoform was identified. Sequence analysis demonstrated the presence of multiple stop codons identifying this isoform of SHC as a processed pseudogene. Using primers designed on the basis of the nucleotide sequence of the pseudogene, we have now amplified and sequenced a human cDNA that encodes the SHC p66 protein. Thus, we have characterized the human SHC p66 isoform cDNA and identified a processed SHC pseudogene that maps to chromosome Xq12- q13.1.

Original languageEnglish
Pages (from-to)349-352
Number of pages4
JournalGenomics
Volume42
Issue number2
DOIs
Publication statusPublished - 1 Jun 1997
Externally publishedYes

Fingerprint

Pseudogenes
Protein Isoforms
Yeast Artificial Chromosomes
Complementary DNA
Chromosomes
Genes
Clone Cells
ras Proteins
Polymerase Chain Reaction
Terminator Codon
3' Untranslated Regions
Fluorescence In Situ Hybridization
Libraries
Sequence Analysis
Cell Differentiation
Proteins
Growth

ASJC Scopus subject areas

  • Genetics

Cite this

Harun, R. B., Smith, K. K., Leek, J. P., Markham, A. F., Morris, A., & Morrison, J. F. J. (1997). Characterization of human SHC p66 cdna and its processed pseudogene mapping to Xq12-q13.1. Genomics, 42(2), 349-352. https://doi.org/10.1006/geno.1997.4728

Characterization of human SHC p66 cdna and its processed pseudogene mapping to Xq12-q13.1. / Harun, Roslan B.; Smith, Kim K.; Leek, Jack P.; Markham, Alexander F.; Morris, Alan; Morrison, John F J.

In: Genomics, Vol. 42, No. 2, 01.06.1997, p. 349-352.

Research output: Contribution to journalArticle

Harun, RB, Smith, KK, Leek, JP, Markham, AF, Morris, A & Morrison, JFJ 1997, 'Characterization of human SHC p66 cdna and its processed pseudogene mapping to Xq12-q13.1', Genomics, vol. 42, no. 2, pp. 349-352. https://doi.org/10.1006/geno.1997.4728
Harun RB, Smith KK, Leek JP, Markham AF, Morris A, Morrison JFJ. Characterization of human SHC p66 cdna and its processed pseudogene mapping to Xq12-q13.1. Genomics. 1997 Jun 1;42(2):349-352. https://doi.org/10.1006/geno.1997.4728
Harun, Roslan B. ; Smith, Kim K. ; Leek, Jack P. ; Markham, Alexander F. ; Morris, Alan ; Morrison, John F J. / Characterization of human SHC p66 cdna and its processed pseudogene mapping to Xq12-q13.1. In: Genomics. 1997 ; Vol. 42, No. 2. pp. 349-352.
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