Identification of hepatocyte growth factor activator inhibitor type 2 (HAI-2)-related small peptide (H2RSP): its nuclear localization and generation of chimeric mRNA transcribed from both HAI-2 and H2RSP genes.
A novel small gene, designated hepatocyte growth factor activator inhibitor type 2 (HAI-2)-related small peptide (H2RSP) was cloned and characterized in the process of the search for splicing variant forms of HAI-2 by 3'-rapid amplification of cDNA ends (RACE). The gene consisted of 4 exons spanning approximately 1 kb and was located in 11 kb downstream of HAI-2 gene (19q.13.11). The novel transcript identified by 3'-RACE was thought to be chimerically transcribed from both HAI-2 (exons 1-7) and H2RSP (exons 2-4) genes. Wild-type H2RSP mRNA (0.5 kb) was detected abundantly in various tissues including the gastrointestinal tract, whereas chimeric mRNA (1.5 kb) was found mainly in the kidney, prostate, and placenta by Northern blot analysis. The predicted amino acid sequence of H2RSP contained two unique domains, namely the serine-rich region (exon 3) and the lysine-rich region (exon 4). Transfection of deleted series of H2RSP cDNAs fused to enhanced green fluorescent protein (EGFP) into HeLa cells revealed that H2RSP has nuclear localization signal in the lysine-rich region. Immunohistochemical study using anti-H2RSP polyclonal antibody indeed revealed the nuclear localization of this peptide in vivo. These results suggest that H2RSP and H2RSP/HAI-2 chimeric peptides might function as a transcriptional regulatory peptide at the nucleus.
Itoh
H
H
Second Department of Pathology, Miyazaki Medical College, 5200 Kihara, Kiyotake, Miyazaki 889-1692, Japan. hitoh@post.miyazaki-med.ac.jp
Kataoka
H
H
Yamauchi
M
M
Naganuma
S
S
Akiyama
Y
Y
Nuki
Y
Y
Shimomura
T
T
Miyazawa
K
K
Kitamura
N
N
Koono
M
M
D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D016466:CHO Cells; D002467:Cell Nucleus / Q000378:metabolism*; D006224:Cricetinae; D018076:DNA, Complementary / Q000032:analysis / Q000302:isolation & purification; D049452:Green Fluorescent Proteins; D006367:HeLa Cells; D017228:Hepatocyte Growth Factor / Q000378:metabolism; D006801:Humans; D007150:Immunohistochemistry; D008164:Luminescent Proteins / Q000378:metabolism; D008562:Membrane Glycoproteins / Q000032:analysis / Q000235:genetics; D008969:Molecular Sequence Data; D009687:Nuclear Proteins / Q000032:analysis / Q000235:genetics*; D012333:RNA, Messenger / Q000378:metabolism; D011993:Recombinant Fusion Proteins / Q000378:metabolism; D012697:Serine Endopeptidases / Q000378:metabolism*; D013347:Subcellular Fractions; D014018:Tissue Distribution; D014157:Transcription Factors / Q000032:analysis / Q000235:genetics*; D014360:Trypsin Inhibitor, Kunitz Soybean
C439765:C19orf33 protein, human; D018076:DNA, Complementary; D008164:Luminescent Proteins; D008562:Membrane Glycoproteins; D009687:Nuclear Proteins; D012333:RNA, Messenger; D011993:Recombinant Fusion Proteins; C109148:SPINT2 protein, human; D014157:Transcription Factors; D049452:Green Fluorescent Proteins; D017228:Hepatocyte Growth Factor; D014360:Trypsin Inhibitor, Kunitz Soybean; C081092:HGF activator; D012697:Serine Endopeptidases