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Analysis of DNA regions directing the tissue-general expression of alpha-1 tubulin in Drosophila melanogaster.

Record Type:	Electronic resources : Monograph/item
Title/Author:	Analysis of DNA regions directing the tissue-general expression of alpha-1 tubulin in Drosophila melanogaster./
Author:	Chen, Chien-Tsu.
Published:	Ann Arbor : ProQuest Dissertations & Theses, : 1993,
Description:	177 p.
Notes:	Source: Dissertations Abstracts International, Volume: 55-02, Section: B.
Contained By:	Dissertations Abstracts International55-02B.
Subject:	Molecular biology. -
Online resource:	http://pqdd.sinica.edu.tw/twdaoapp/servlet/advanced?query=9322344
ISBN:	9798208526835

Analysis of DNA regions directing the tissue-general expression of alpha-1 tubulin in Drosophila melanogaster.
Chen, Chien-Tsu.

Analysis of DNA regions directing the tissue-general expression of alpha-1 tubulin in Drosophila melanogaster. - Ann Arbor : ProQuest Dissertations & Theses, 1993 - 177 p.

Source: Dissertations Abstracts International, Volume: 55-02, Section: B.

Thesis (Ph.D.)--Brandeis University, 1993.

This item must not be sold to any third party vendors.

To examine the expression of the Drosophila melanogaster alpha-1 tubulin gene, $T\\alpha$1, a major portion of the gene ($-$2000 to $+$696 bp) was fused to the E. coli lacZ gene and the resulting chimera introduced into Drosophila germline by P-element transformation. All transgenic lines showed a high level of $\\beta$-galactosidase activity at all developmental stages and in all living tissues when assayed at the dissecting microscope level of resolution using a histological staining detection method. An appropriate genetic cross demonstrated that the paternal copy of the $T\\alpha$1 fusion gene is first expressed at embryonic stage 5. The term tissue-general expression is adopted to describe this $T\\alpha$1 expression pattern. Localization of the transcriptional regulators in the $T\\alpha$1 DNA by deletion and substitution mutations revealed that two regulators, UR ($-$157 to $-$34 bp) and DR ($+$20 to $+$696 bp), are necessary and together are sufficient to direct tissue-general expression in vivo from the $T\\alpha$1 promoter ($-$34 to $+$20 bp). These studies also showed that the promoter of the Hsp70 gene ($-$44 to $+$35 bp) can also receive the $T\\alpha$1 regulatory signals to give tissue-general expression, indicating that the $T\\alpha$1 promoter is not uniquely adapted to be regulated by UR and DR. The arrangement of UR and DR relative to the promoter was shown to be important for tissue-general expression, indicating that these regulators are not simple enhancers. Tests of promoter specificity in receiving tissue-general regulation from UR and DR demonstrated that both tissue-specific and tissue-general promoters can be regulated to express tissue-generally, but that the Hsp70 promoter has unique properties. In vitro transcription assays revealed that both the tissue-general, inducible promoter from Hsp70 and several tissue-specific promoters (Yp1, adh distal, and Act5C distal), are regulated in vitro by UR. Germline transformation experiments showed that these three normally tissue-specific promoters can also receive UR regulation in vivo, resulting, when combined with the influence of DR, in tissue-general expression. By contrast, the Hsp70 promoter requires only DR and not UR for tissue-general expression in vivo. Sequence comparisons suggest that only the TATA sequence is necessary to receive the tissue-general regulation from the combination of UR and DR. Further, the sequence comparisons indicate that the Hsp70 promoter may contain regulators that substitute for UR activity. The results also indicate that UR and DR block chromosomal position effects on the transcription of promoters from both tissue-general and tissue-specific genes.

ISBN: 9798208526835Subjects--Topical Terms:

517296
Molecular biology.
Subjects--Index Terms:

alpha-1 tubulin

Analysis of DNA regions directing the tissue-general expression of alpha-1 tubulin in Drosophila melanogaster.
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245 1 0 $a Analysis of DNA regions directing the tissue-general expression of alpha-1 tubulin in Drosophila melanogaster.
260 1 $a Ann Arbor : $b ProQuest Dissertations & Theses, $c 1993
300 $a 177 p.
500 $a Source: Dissertations Abstracts International, Volume: 55-02, Section: B.
500 $a Publisher info.: Dissertation/Thesis.
500 $a Advisor: Wensink, Pieter C.
502 $a Thesis (Ph.D.)--Brandeis University, 1993.
506 $a This item must not be sold to any third party vendors.
506 $a This item must not be added to any third party search indexes.
520 $a To examine the expression of the Drosophila melanogaster alpha-1 tubulin gene, $T\\alpha$1, a major portion of the gene ($-$2000 to $+$696 bp) was fused to the E. coli lacZ gene and the resulting chimera introduced into Drosophila germline by P-element transformation. All transgenic lines showed a high level of $\\beta$-galactosidase activity at all developmental stages and in all living tissues when assayed at the dissecting microscope level of resolution using a histological staining detection method. An appropriate genetic cross demonstrated that the paternal copy of the $T\\alpha$1 fusion gene is first expressed at embryonic stage 5. The term tissue-general expression is adopted to describe this $T\\alpha$1 expression pattern. Localization of the transcriptional regulators in the $T\\alpha$1 DNA by deletion and substitution mutations revealed that two regulators, UR ($-$157 to $-$34 bp) and DR ($+$20 to $+$696 bp), are necessary and together are sufficient to direct tissue-general expression in vivo from the $T\\alpha$1 promoter ($-$34 to $+$20 bp). These studies also showed that the promoter of the Hsp70 gene ($-$44 to $+$35 bp) can also receive the $T\\alpha$1 regulatory signals to give tissue-general expression, indicating that the $T\\alpha$1 promoter is not uniquely adapted to be regulated by UR and DR. The arrangement of UR and DR relative to the promoter was shown to be important for tissue-general expression, indicating that these regulators are not simple enhancers. Tests of promoter specificity in receiving tissue-general regulation from UR and DR demonstrated that both tissue-specific and tissue-general promoters can be regulated to express tissue-generally, but that the Hsp70 promoter has unique properties. In vitro transcription assays revealed that both the tissue-general, inducible promoter from Hsp70 and several tissue-specific promoters (Yp1, adh distal, and Act5C distal), are regulated in vitro by UR. Germline transformation experiments showed that these three normally tissue-specific promoters can also receive UR regulation in vivo, resulting, when combined with the influence of DR, in tissue-general expression. By contrast, the Hsp70 promoter requires only DR and not UR for tissue-general expression in vivo. Sequence comparisons suggest that only the TATA sequence is necessary to receive the tissue-general regulation from the combination of UR and DR. Further, the sequence comparisons indicate that the Hsp70 promoter may contain regulators that substitute for UR activity. The results also indicate that UR and DR block chromosomal position effects on the transcription of promoters from both tissue-general and tissue-specific genes.
590 $a School code: 0021.
650 4 $a Molecular biology. $3 517296
650 4 $a Cellular biology. $3 3172791
650 4 $a Genetics. $3 530508
653 $a alpha-1 tubulin
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710 2 $a Brandeis University. $3 1017415
773 0 $t Dissertations Abstracts International $g 55-02B.
790 $a 0021
791 $a Ph.D.
792 $a 1993
793 $a English
856 4 0 $u http://pqdd.sinica.edu.tw/twdaoapp/servlet/advanced?query=9322344