Molecular analysis of the GCF gene identifies revisions to the cDNA and amino acid sequences(1).
GC factor (GCF) was reported as a transcriptional regulator that binds to a specific GC-rich sequence in the epidermal growth factor receptor (EGFR) gene promoter and represses its expression. In this paper, we present the data on three revisions of the cDNA sequence that lead to significant changes of the amino acid sequences of the published GCF. Firstly, 5'-rapid amplification of cDNA end (5'-RACE) analysis revealed that the 308 nucleotides of 5'-end of the previously published GCF cDNA does not exist at the 5'-end of the RACE product. Simultaneously, the correct 5'-end cDNA sequence of 31 nucleotides was identified. Secondly, the 'T' at the position 787 of the published GCF cDNA was not observed. Finally, a new sequence of 114 nucleotides was identified between the positions 851 and 852 of the published cDNA sequence. The revisions result in a GCF cDNA of 2661 nucleotides that encodes a protein of 781 amino acids, replacing the highly basic region of the amino-terminus of the published GCF with a new sequence of 147 amino acids. In this era of massive gene cloning and sequencing, this study is a warning to the biological research of recent years.
Takimoto
M
M
Division of Gene Regulation, Cancer Institute, Hokkaido University, School of Medicine, N15, W7, Kita-Ku, Sapporo, Hokkaido, Japan. takimoto@med.hokudai.ac.jp
Mao
P
P
Wei
G
G
Yamazaki
H
H
Miura
T
T
Johnson
A C
AC
Kuzumaki
N
N
D017398:Alternative Splicing; D000595:Amino Acid Sequence; D018076:DNA, Complementary / Q000737:chemistry; D006801:Humans; D007962:Leukocytes / Q000378:metabolism; D008969:Molecular Sequence Data; D012333:RNA, Messenger / Q000737:chemistry; D012097:Repressor Proteins / Q000737:chemistry / Q000235:genetics*; D020133:Reverse Transcriptase Polymerase Chain Reaction; D014407:Tumor Cells, Cultured
D018076:DNA, Complementary; C072278:GCFC2 protein, human; D012333:RNA, Messenger; D012097:Repressor Proteins