NEWLY SYNTHESIZED RNA - SIMULTANEOUS MEASUREMENT IN INTACT-CELLS OF TRANSCRIPTION RATES AND RNA STABILITY OF INSULIN-LIKE GROWTH FACTOR-I, ACTIN, AND ALBUMIN IN GROWTH HORMONE-STIMULATED HEPATOCYTES

被引:64
作者
JOHNSON, TR [1 ]
RUDIN, SD [1 ]
BLOSSEY, BK [1 ]
ILAN, J [1 ]
ILAN, J [1 ]
机构
[1] CASE WESTERN RESERVE UNIV,SCH MED,FACHBEREICH REPROD BIOL,CLEVELAND,OH 44106
关键词
4-THIOURIDINE; CULTURED HEPATOCYTES; MERCURATED AGAROSE AFFINITY CHROMATOGRAPHY;
D O I
10.1073/pnas.88.12.5287
中图分类号
O [数理科学和化学]; P [天文学、地球科学]; Q [生物科学]; N [自然科学总论];
学科分类号
07 ; 0710 ; 09 ;
摘要
The levels of several RNA transcripts in cultured hepatocytes are regulated by transcriptional and post-transcriptional mechanisms and are affected by growth hormone and insulin. We assessed the effects of these hormones on transcription rates and the stability of insulin-like growth factor I, actin, and albumin transcripts in intact cells of primary cultures of rat hepatocytes by analyzing thiol-labeled, newly synthesized RNA isolated by mercurated agarose affinity chromatography. The application of this concept to the measurement of transcript stability is presented in detail. The data indicate that growth hormone stimulates the transcription rates of insulin-like growth factor I, actin, and albumin genes. The stability of all three transcripts, particularly albumin, appears to be lower in growth hormone-containing medium than it is in insulin-containing medium. The experiments indicate that the rates of transcription and/or degradation of albumin mRNA are influenced by hormonal treatment. However, the cells maintain roughly constant albumin transcript levels independent of hormone treatment by compensatory changes in the rates of transcription and degradation.
引用
收藏
页码:5287 / 5291
页数:5
相关论文
共 20 条
[1]  
AIELLO LP, 1988, J BIOL CHEM, V263, P4013
[2]  
BEZZARA R, 1990, BIOCHEM BIOPH RES CO, V166, P29
[3]   INDUCTION AND DECAY OF HUMAN FIBROBLAST INTERFERON MESSENGER-RNA [J].
CAVALIERI, RL ;
HAVELL, EA ;
VILCEK, J ;
PESTKA, S .
PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA, 1977, 74 (10) :4415-4419
[4]  
CHOMCZYNSKI P, 1987, ANAL BIOCHEM, V162, P156, DOI 10.1016/0003-2697(87)90021-2
[5]  
CRAMER CL, 1985, SCIENCE, V227, P1240
[6]   PROLACTIN-MEDIATED TRANSCRIPTIONAL AND POST-TRANSCRIPTIONAL CONTROL OF CASEIN GENE-EXPRESSION [J].
GUYETTE, WA ;
MATUSIK, RJ ;
ROSEN, JM .
CELL, 1979, 17 (04) :1013-1023
[7]   CHARACTERIZATION OF AN INDUCIBLE PROMOTER SYSTEM TO INVESTIGATE DECAY OF STABLE MESSENGER-RNA MOLECULES [J].
HELMS, SR ;
ROTTMAN, FM .
NUCLEIC ACIDS RESEARCH, 1990, 18 (02) :255-259
[8]   EXPRESSION FROM THE TRANSFERRIN GENE PROMOTER IN TRANSGENIC MICE [J].
IDZERDA, RL ;
BEHRINGER, RR ;
THEISEN, M ;
HUGGENVIK, JI ;
MCKNIGHT, GS ;
BRINSTER, RL .
MOLECULAR AND CELLULAR BIOLOGY, 1989, 9 (11) :5154-5162
[9]   MAINTENANCE OF DIFFERENTIATED RAT HEPATOCYTES IN PRIMARY CULTURE [J].
ISOM, HC ;
SECOTT, T ;
GEORGOFF, I ;
WOODWORTH, C ;
MUMMAW, J .
PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA, 1985, 82 (10) :3252-3256
[10]  
JOHNSON T R, 1990, Journal of Cell Biology, V111, p505A