Dual functions of a cis-acting element within the rat prolactin gene promoter

1989 ◽  
Vol 9 (2) ◽  
pp. 817-819
Author(s):  
E A Barron ◽  
Z Cao ◽  
B G Schneider ◽  
E Kraig ◽  
A J Carrillo ◽  
...  
Keyword(s):  
Promoter Region ◽  
Gene Promoter ◽  
In Vitro Transcription ◽  
Cis Acting ◽  
Transcription System ◽  
Prolactin Gene ◽  
Imperfect Palindrome ◽  
Dual Functions ◽  
Symmetrical Sequence

Within the promoter region of the rat prolactin gene lies a TA-rich imperfect palindrome. The possible functions of the 18-base-pair symmetrical sequence were investigated by using an in vitro transcription system. Prolactin templates with and without the palindrome were transcriptionally assayed in both pituitary and nonpituitary extracts. Our results indicated that the palindromic sequence has at least two functions in the regulation of prolactin transcription.

scholarly journals Dual functions of a cis-acting element within the rat prolactin gene promoter.

1989 ◽  
Vol 9 (2) ◽  
pp. 817-819 ◽  
Author(s):  
E A Barron ◽  
Z Cao ◽  
B G Schneider ◽  
E Kraig ◽  
A J Carrillo ◽  
...  
Keyword(s):  
Promoter Region ◽  
Gene Promoter ◽  
In Vitro Transcription ◽  
Cis Acting ◽  
Transcription System ◽  
Prolactin Gene ◽  
Imperfect Palindrome ◽  
Dual Functions ◽  
Symmetrical Sequence

Within the promoter region of the rat prolactin gene lies a TA-rich imperfect palindrome. The possible functions of the 18-base-pair symmetrical sequence were investigated by using an in vitro transcription system. Prolactin templates with and without the palindrome were transcriptionally assayed in both pituitary and nonpituitary extracts. Our results indicated that the palindromic sequence has at least two functions in the regulation of prolactin transcription.

Architecture of the maize mitochondrial atp1 promoter as determined by linker-scanning and point mutagenesis

1993 ◽  
Vol 13 (12) ◽  
pp. 7232-7238
Author(s):  
W D Rapp ◽  
D S Lupold ◽  
S Mack ◽  
D B Stern
Keyword(s):  
Promoter Region ◽  
Consensus Sequence ◽  
Point Mutations ◽  
In Vitro Transcription ◽  
Wild Type ◽  
Transcription Start ◽  
Transcription System ◽  
Mitochondrial Promoters ◽  
Point Mutagenesis

Plant mitochondrial promoters are poorly conserved but generally share a loose consensus sequence spanning approximately 17 nucleotides. Using a homologous in vitro transcription system, we have previously shown that an 11-nucleotide sequence within this region comprises at least part of the maize mitochondrial atp1 promoter (W. Rapp and D. Stern, EMBO J. 11:1065-1073, 1992). We have extended this finding by using a series of linker-scanning and point mutations to define the atp1 promoter in detail. Our results show that mutations at positions -12 to +5, relative to the major transcription start site, can decrease initiation rates to between < 10 and 40% of wild-type levels. Some mutations, scattered throughout this region, have lesser effects or no effect. Taken together, our data suggest a model in which the atp1 promoter consists of a central domain extending from -7 to +5 and an upstream domain of 1 to 3 bp that is centered around -11 to -12. Because many mutations within this promoter region are tolerated in vitro, the maize atp1 promoter is distinct from the highly conserved yeast mitochondrial promoters.

scholarly journals A Positive cis-Acting DNA Element Is Required for High-Level Transcription in Chlamydia

2000 ◽  
Vol 182 (18) ◽  
pp. 5167-5171 ◽  
Author(s):  
Chris S. Schaumburg ◽  
Ming Tan
Keyword(s):  
Escherichia Coli ◽  
Chlamydia Trachomatis ◽  
Rna Polymerase ◽  
Promoter Region ◽  
In Vitro Transcription ◽  
Transcription Assay ◽  
Cis Acting ◽  
E Coli ◽  
High Level

ABSTRACT The spacer A/T region is a positive cis-acting DNA element that was identified in the Chlamydia trachomatisrRNA promoter region. We have now demonstrated that similar sequences in other chlamydial promoters are important for transcription. Substitution of candidate spacer A/T regions in four chlamydial promoters decreased transcription by partially purified C. trachomatis RNA polymerase in an in vitro transcription assay. Addition of a spacer A/T region to the dnaK promoter, which does not contain an identifiable spacer A/T region, increased transcription 16-fold. Transcription of Escherichia colipromoters by C. trachomatis RNA polymerase also appeared to be dependent on the spacer A/T region. However, the effect of the spacer A/T region on transcription by E. coli RNA polymerase was small. In summary, the spacer A/T region is a novel DNA element that is required for high-level transcription of many promoters by chlamydial RNA polymerase.

scholarly journals Reconstitution of cell-type-specific transcription of the rat prolactin gene in vitro.

1987 ◽  
Vol 7 (10) ◽  
pp. 3402-3408 ◽  
Author(s):  
Z D Cao ◽  
E A Barron ◽  
A J Carillo ◽  
Z D Sharp
Keyword(s):  
Gene Expression ◽  
Rna Polymerase Ii ◽  
Expression System ◽  
In Vitro Transcription ◽  
Gh3 Cells ◽  
Nuclear Extracts ◽  
Prolactin Gene ◽  
Flanking Region ◽  
Imperfect Palindrome

We present evidence for the existence of prolactin upstream factor 1 (PUF-1) in rat pituitary-derived cells and demonstrate its interaction with a symmetrical DNA element located in the 5' flanking region of the gene. An in vitro expression system developed from pituitary-derived GH3 cells was used to determine that 420 base pairs (bp) of 5' flanking DNA was sufficient for cell-specific, accurate, and efficient RNA polymerase II transcription of the rat prolactin gene. Reconstitution of in vitro transcription with pituitary and nonpituitary nuclear extracts suggested that the presence of GH3 cell-specific factors mediated the activation of prolactin gene expression. We also demonstrated that a functionally stable transcription complex assembled on the prolactin promoter. Using DNase I protection procedures, we have identified the DNA-protein binding area in the prolactin 5' flanking region. GH3 nuclear extracts contain a cell-specific protein (PUF-I) that binds to a 28-bp region (-63 to -36)which contains an 18-bp imperfect palindrome (-63 to -46). The role that the interaction between PUF-I and the imperfect palindrome plays in in vitro pituitary-specific prolactin gene expression is discussed.

scholarly journals Determination of the promoter region of mouse ribosomal RNA gene by an in vitro transcription system.

1984 ◽  
Vol 81 (2) ◽  
pp. 299-303 ◽  
Author(s):  
O. Yamamoto ◽  
N. Takakusa ◽  
Y. Mishima ◽  
R. Kominami ◽  
M. Muramatsu
Keyword(s):  
Ribosomal Rna ◽  
Promoter Region ◽  
In Vitro Transcription ◽  
Ribosomal Rna Gene ◽  
Transcription System

scholarly journals Architecture of the maize mitochondrial atp1 promoter as determined by linker-scanning and point mutagenesis.

1993 ◽  
Vol 13 (12) ◽  
pp. 7232-7238 ◽  
Author(s):  
W D Rapp ◽  
D S Lupold ◽  
S Mack ◽  
D B Stern
Keyword(s):  
Promoter Region ◽  
Consensus Sequence ◽  
Point Mutations ◽  
In Vitro Transcription ◽  
Wild Type ◽  
Transcription Start ◽  
Transcription System ◽  
Mitochondrial Promoters ◽  
Point Mutagenesis

Plant mitochondrial promoters are poorly conserved but generally share a loose consensus sequence spanning approximately 17 nucleotides. Using a homologous in vitro transcription system, we have previously shown that an 11-nucleotide sequence within this region comprises at least part of the maize mitochondrial atp1 promoter (W. Rapp and D. Stern, EMBO J. 11:1065-1073, 1992). We have extended this finding by using a series of linker-scanning and point mutations to define the atp1 promoter in detail. Our results show that mutations at positions -12 to +5, relative to the major transcription start site, can decrease initiation rates to between < 10 and 40% of wild-type levels. Some mutations, scattered throughout this region, have lesser effects or no effect. Taken together, our data suggest a model in which the atp1 promoter consists of a central domain extending from -7 to +5 and an upstream domain of 1 to 3 bp that is centered around -11 to -12. Because many mutations within this promoter region are tolerated in vitro, the maize atp1 promoter is distinct from the highly conserved yeast mitochondrial promoters.

Reconstitution of cell-type-specific transcription of the rat prolactin gene in vitro

1987 ◽  
Vol 7 (10) ◽  
pp. 3402-3408
Author(s):  
Z D Cao ◽  
E A Barron ◽  
A J Carillo ◽  
Z D Sharp
Keyword(s):  
Gene Expression ◽  
Rna Polymerase Ii ◽  
Expression System ◽  
In Vitro Transcription ◽  
Gh3 Cells ◽  
Nuclear Extracts ◽  
Prolactin Gene ◽  
Flanking Region ◽  
Imperfect Palindrome

We present evidence for the existence of prolactin upstream factor 1 (PUF-1) in rat pituitary-derived cells and demonstrate its interaction with a symmetrical DNA element located in the 5' flanking region of the gene. An in vitro expression system developed from pituitary-derived GH3 cells was used to determine that 420 base pairs (bp) of 5' flanking DNA was sufficient for cell-specific, accurate, and efficient RNA polymerase II transcription of the rat prolactin gene. Reconstitution of in vitro transcription with pituitary and nonpituitary nuclear extracts suggested that the presence of GH3 cell-specific factors mediated the activation of prolactin gene expression. We also demonstrated that a functionally stable transcription complex assembled on the prolactin promoter. Using DNase I protection procedures, we have identified the DNA-protein binding area in the prolactin 5' flanking region. GH3 nuclear extracts contain a cell-specific protein (PUF-I) that binds to a 28-bp region (-63 to -36)which contains an 18-bp imperfect palindrome (-63 to -46). The role that the interaction between PUF-I and the imperfect palindrome plays in in vitro pituitary-specific prolactin gene expression is discussed.

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