Identification of G protein alpha subunit mutations in human growth hormone (GH)- and GH/prolactin-secreting pituitary tumors by single-strand conformation polymorphism (SSCP) analysis.
We have applied the polymerase chain reaction (PCR) and single-strand conformation polymorphism analysis (SSCP) to detect activating mutations in the Gs alpha subunit gene, amplifying genomic DNA extracted from growth hormone (GH)- and GH/prolactin (PRL)-secreting human pituitary tumors. Of 15 tumors tested six contained mutations in the analyzed regions of the Gs alpha. SSCP analysis revealed band shift in exon 8 in four GH- and in one GH/PRL-secreting tumors, and in exon 9 in one GH/PRL-secreting tumor. Direct sequencing of PCR reaction products identified the mutations as R201-H, R201-S and R201-C in exon 8 and Q227-L in exon 9. These results show the efficacy of PCR/SSCP analysis in the detection of G protein mutations and extend the generalization that these sites are hot spots in tumor-inducing mutations.
Drews
R T
RT
Research Unit on Biology of Reproduction and Development, Ste-Justine Hospital, Université de Montréal, Canada.
Gravel
R A
RA
Collu
R
R
D000236:Adenoma / Q000235:genetics* / Q000378:metabolism; D001483:Base Sequence; D003062:Codon; D004252:DNA Mutational Analysis; D004273:DNA, Neoplasm / Q000235:genetics*; D004277:DNA, Single-Stranded / Q000235:genetics*; D004789:Enzyme Activation; D005091:Exons; D020558:GTP Phosphohydrolases / Q000235:genetics*; D019204:GTP-Binding Proteins / Q000235:genetics*; D013006:Growth Hormone / Q000378:metabolism*; D006801:Humans; D008969:Molecular Sequence Data; D009154:Mutation; D009363:Neoplasm Proteins / Q000235:genetics* / Q000378:metabolism; D009690:Nucleic Acid Conformation; D010911:Pituitary Neoplasms / Q000235:genetics* / Q000378:metabolism; D016133:Polymerase Chain Reaction; D011388:Prolactin / Q000378:metabolism*
D003062:Codon; D004273:DNA, Neoplasm; D004277:DNA, Single-Stranded; D009363:Neoplasm Proteins; D011388:Prolactin; D013006:Growth Hormone; D020558:GTP Phosphohydrolases; D019204:GTP-Binding Proteins