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A genetic analysis of the insulin tr...

Mamounas, Michael Joseph,

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A genetic analysis of the insulin transmembrane mitogenic signal in Chinese hamster ovary cells /

Record Type:	Electronic resources : Monograph/item
Title/Author:	A genetic analysis of the insulin transmembrane mitogenic signal in Chinese hamster ovary cells // Michael Joseph Mamounas.
Author:	Mamounas, Michael Joseph,
Description:	1 electronic resource (173 pages)
Notes:	Source: Dissertations Abstracts International, Volume: 54-02, Section: B.
Contained By:	Dissertations Abstracts International54-02B.
Subject:	Genetics. -
Online resource:	https://pqdd.sinica.edu.tw/twdaoapp/servlet/advanced?query=9218461
ISBN:	9798207272160

A genetic analysis of the insulin transmembrane mitogenic signal in Chinese hamster ovary cells /
Mamounas, Michael Joseph,

A genetic analysis of the insulin transmembrane mitogenic signal in Chinese hamster ovary cells /Michael Joseph Mamounas. - 1 electronic resource (173 pages)

Source: Dissertations Abstracts International, Volume: 54-02, Section: B.

Insulin is the only hormone required for continued growth of Chinese hamster ovary (CHO-K1) cells in the defined medium M-F12. When CHO-K1 cells are incubated in M-F12 without insulin for 48-72 hr, the cells accumulate in Go/G1. In response to physiological concentrations of insulin an 18-fold increase in rate of DNA synthesis occurs due to cells entering S phase after an 8- to 10-hr lag, followed by cell division after 24 hr. The observed mitogenic effect of insulin is due to insulin binding to its own receptor. Drug inhibition studies indicate that RNA synthesis is required for progression to S phase. Insulin-independent mutants can be isolated which have a rate of DNA synthesis and cell division in the absence of insulin comparable to the WT growing with insulin. Insulin binding to its receptor stimulates an increase in abundance of mRNA for c-fos, c-jun, Krox-20, Krox-24, fra-1, jun-B, c-myc, and JE. The mutants have constitutive levels of Krox-24, fra-1, jun-B, c-myc and JE that are dependent on the time in culture and/or the cell density. One mutant, I$\\sp{\\rm in}$3, appears to regulate these genes by an autocrine mechanism. There were no detectable levels of mRNA for genes c-fos and Krox-20 and no increase in level of mRNA for c-jun (INS-type 1 genes) in the unstimulated mutants. The increase in abundance of fra-1, jun-B, and possibly c-myc, in both the induced WT and the uninduced mutant 27, can be accounted for by increased rates of transcription defining a class of genes referred to as INS-type 2$\\alpha$. In contrast, JE is regulated by a post-transcriptional mechanism defining a class of genes referred to as INS-type 2$\\beta$. Although these INS-type 1 and type 2 genes may be involved in the full insulin-mediated mitogenic signal, the constitutive up-regulation of only the INS-type 2 genes is sufficient for insulin-independent growth. Analysis of hybrids constructed between WT and mutant 27 indicates that the mutant phenotype is recessive, pointing to the existence of a regulatory gene producing a negative regulator which controls the expression of both INS-type 2 genes and DNA synthesis.

English

ISBN: 9798207272160Subjects--Topical Terms:

530508
Genetics.

A genetic analysis of the insulin transmembrane mitogenic signal in Chinese hamster ovary cells /
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100 1 $a Mamounas, Michael Joseph, $e author. $3 3825658
245 1 2 $a A genetic analysis of the insulin transmembrane mitogenic signal in Chinese hamster ovary cells / $c Michael Joseph Mamounas.
264 1 $a Ann Arbor : $b ProQuest Dissertations & Theses, $c 1991
300 $a 1 electronic resource (173 pages)
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500 $a Source: Dissertations Abstracts International, Volume: 54-02, Section: B.
500 $a Publisher info.: Dissertation/Thesis.
500 $a Advisors: Englesberg, Ellis.
502 $b Ph.D. $c University of California, Santa Barbara $d 1991.
520 $a Insulin is the only hormone required for continued growth of Chinese hamster ovary (CHO-K1) cells in the defined medium M-F12. When CHO-K1 cells are incubated in M-F12 without insulin for 48-72 hr, the cells accumulate in Go/G1. In response to physiological concentrations of insulin an 18-fold increase in rate of DNA synthesis occurs due to cells entering S phase after an 8- to 10-hr lag, followed by cell division after 24 hr. The observed mitogenic effect of insulin is due to insulin binding to its own receptor. Drug inhibition studies indicate that RNA synthesis is required for progression to S phase. Insulin-independent mutants can be isolated which have a rate of DNA synthesis and cell division in the absence of insulin comparable to the WT growing with insulin. Insulin binding to its receptor stimulates an increase in abundance of mRNA for c-fos, c-jun, Krox-20, Krox-24, fra-1, jun-B, c-myc, and JE. The mutants have constitutive levels of Krox-24, fra-1, jun-B, c-myc and JE that are dependent on the time in culture and/or the cell density. One mutant, I$\\sp{\\rm in}$3, appears to regulate these genes by an autocrine mechanism. There were no detectable levels of mRNA for genes c-fos and Krox-20 and no increase in level of mRNA for c-jun (INS-type 1 genes) in the unstimulated mutants. The increase in abundance of fra-1, jun-B, and possibly c-myc, in both the induced WT and the uninduced mutant 27, can be accounted for by increased rates of transcription defining a class of genes referred to as INS-type 2$\\alpha$. In contrast, JE is regulated by a post-transcriptional mechanism defining a class of genes referred to as INS-type 2$\\beta$. Although these INS-type 1 and type 2 genes may be involved in the full insulin-mediated mitogenic signal, the constitutive up-regulation of only the INS-type 2 genes is sufficient for insulin-independent growth. Analysis of hybrids constructed between WT and mutant 27 indicates that the mutant phenotype is recessive, pointing to the existence of a regulatory gene producing a negative regulator which controls the expression of both INS-type 2 genes and DNA synthesis.
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650 4 $a Genetics. $3 530508
650 4 $a Cellular biology. $3 3172791
650 4 $a Molecular biology. $3 517296
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710 2 $a University of California, Santa Barbara. $e degree granting institution. $3 3818149
720 1 $a Englesberg, Ellis $e degree supervisor.
773 0 $t Dissertations Abstracts International $g 54-02B.
790 $a 0035
791 $a Ph.D.
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856 4 0 $u https://pqdd.sinica.edu.tw/twdaoapp/servlet/advanced?query=9218461