Effects of procollagen C-proteinase enhancer protein on the growth of cultured rat fibroblasts revealed by an excisable retroviral vector.
An excisable retroviral vector, TSN-lox, was developed by exploiting Cre-loxP homologous recombination. An integrated TSN-lox provirus could be excised, leaving a solo long terminal repeat at the integration site; inverse PCR, taking advantage of the solo long terminal repeat, was used to characterize cellular flanking sequences. A TSN-lox-transduced Rat2 cell clone, lox-7, was found to harbor the provirus in an intron of the procollagen C-proteinase enhancer protein (PCPE) gene, whose expression was lowered compared with that of the parental Rat2. When the vector provirus in lox-7 cells was excised, PCPE expression was elevated. The level of PCPE expression seemed to affect cell growth properties such as morphology, contact inhibition, and anchorage-independent growth. These results suggested that the excisable retroviral vector may be useful for studying the molecular basis for proviral insertion mutagenesis, and that PCPE may play a significant role in controlling cell growth and differentiation.
Masuda
M
M
Department of Microbiology, Graduate School of Medicine, The University of Tokyo, Japan. mmasuda@m.u-tokyo.ac.jp
Igarashi
H
H
Kano
M
M
Yoshikura
H
H
D000818:Animals; D001483:Base Sequence; D001665:Binding Sites; D015152:Blotting, Northern; D002448:Cell Adhesion / Q000235:genetics; D002455:Cell Division / Q000235:genetics; D002460:Cell Line; D048429:Cell Size / Q000235:genetics; D005347:Fibroblasts / Q000166:cytology; D005786:Gene Expression Regulation; D005822:Genetic Vectors; D006023:Glycoproteins / Q000096:biosynthesis* / Q000235:genetics; D019426:Integrases; D036341:Intercellular Signaling Peptides and Proteins; D007438:Introns; D008969:Molecular Sequence Data; D011533:Proviruses / Q000235:genetics; D012333:RNA, Messenger / Q000032:analysis; D051381:Rats; D012091:Repetitive Sequences, Nucleic Acid; D012190:Retroviridae; D014162:Transfection / Q000379:methods*; D014764:Viral Proteins; D016662:Virus Integration
D006023:Glycoproteins; D036341:Intercellular Signaling Peptides and Proteins; C491554:Pcolce protein, rat; D012333:RNA, Messenger; D014764:Viral Proteins; C045073:Cre recombinase; D019426:Integrases