Functional analysis of the endoxylanase B (xynB) promoter from Penicillium purpurogenum

Jheimmy Díaz, Renato Chávez, Luis F. Larrondo, Jaime Eyzaguirre, Paulina Bull

Resultado de la investigación: Article

5 Citas (Scopus)

Resumen

In Penicillium purpurogenum, the gene encoding endoxylanase B (xynB) is highly expressed by xylan and repressed by glucose at the transcriptional level. The promoter of this gene has a modular structure, with eight putative XlnR binding sites in tandem (XlnR module), and upstream from them, four putative CreA binding sites (CreA module). Promoter fragments containing different modules were inserted into a plasmid, pAN49-1, which contains a basal fungal promoter linked to a reporter gene (lacZ) and its expression was studied in vivo in Aspergillus nidulans. The XlnR module is able to trigger high β-galactosidase activity in the presence of xylan, but the lack of most XlnR sites notoriously reduces this enzymatic activity. No enzyme induction is observed if the orientation of the promoter fragment is inverted. The presence of the CreA module is necessary for glucose repression when β-galactosidase activity is previously induced by xylan. However, when transformant strains containing the XlnR module but lacking all CreA sites were grown in glucose without pre-induction in xylan, a low β-galactosidase activity was observed compared with the same transformants grown in xylan. These results agree with a double-lock regulatory mechanism for both direct and indirect repression of xylanolytic genes by glucose.

Idioma originalEnglish
Páginas (desde-hasta)133-141
Número de páginas9
PublicaciónCurrent Genetics
Volumen54
N.º3
DOI
EstadoPublished - 28 jul 2008

Huella dactilar

Endo-1,4-beta Xylanases
Xylans
Penicillium
Galactosidases
Glucose
Binding Sites
Genes
Aspergillus nidulans
Enzyme Induction
Reporter Genes
Plasmids

ASJC Scopus subject areas

  • Genetics

Citar esto

Díaz, Jheimmy ; Chávez, Renato ; Larrondo, Luis F. ; Eyzaguirre, Jaime ; Bull, Paulina. / Functional analysis of the endoxylanase B (xynB) promoter from Penicillium purpurogenum. En: Current Genetics. 2008 ; Vol. 54, N.º 3. pp. 133-141.
@article{a330a2b627734564be617620cc52f824,
title = "Functional analysis of the endoxylanase B (xynB) promoter from Penicillium purpurogenum",
abstract = "In Penicillium purpurogenum, the gene encoding endoxylanase B (xynB) is highly expressed by xylan and repressed by glucose at the transcriptional level. The promoter of this gene has a modular structure, with eight putative XlnR binding sites in tandem (XlnR module), and upstream from them, four putative CreA binding sites (CreA module). Promoter fragments containing different modules were inserted into a plasmid, pAN49-1, which contains a basal fungal promoter linked to a reporter gene (lacZ) and its expression was studied in vivo in Aspergillus nidulans. The XlnR module is able to trigger high β-galactosidase activity in the presence of xylan, but the lack of most XlnR sites notoriously reduces this enzymatic activity. No enzyme induction is observed if the orientation of the promoter fragment is inverted. The presence of the CreA module is necessary for glucose repression when β-galactosidase activity is previously induced by xylan. However, when transformant strains containing the XlnR module but lacking all CreA sites were grown in glucose without pre-induction in xylan, a low β-galactosidase activity was observed compared with the same transformants grown in xylan. These results agree with a double-lock regulatory mechanism for both direct and indirect repression of xylanolytic genes by glucose.",
keywords = "Aspergillus nidulans, CreA sites, In vivo expression, Penicillium purpurogenum, Promoter fragments, XlnR sites",
author = "Jheimmy D{\'i}az and Renato Ch{\'a}vez and Larrondo, {Luis F.} and Jaime Eyzaguirre and Paulina Bull",
year = "2008",
month = "7",
day = "28",
doi = "10.1007/s00294-008-0205-y",
language = "English",
volume = "54",
pages = "133--141",
journal = "Current Genetics",
issn = "0172-8083",
publisher = "Springer Verlag",
number = "3",

}

Functional analysis of the endoxylanase B (xynB) promoter from Penicillium purpurogenum. / Díaz, Jheimmy; Chávez, Renato; Larrondo, Luis F.; Eyzaguirre, Jaime; Bull, Paulina.

En: Current Genetics, Vol. 54, N.º 3, 28.07.2008, p. 133-141.

Resultado de la investigación: Article

TY - JOUR

T1 - Functional analysis of the endoxylanase B (xynB) promoter from Penicillium purpurogenum

AU - Díaz, Jheimmy

AU - Chávez, Renato

AU - Larrondo, Luis F.

AU - Eyzaguirre, Jaime

AU - Bull, Paulina

PY - 2008/7/28

Y1 - 2008/7/28

N2 - In Penicillium purpurogenum, the gene encoding endoxylanase B (xynB) is highly expressed by xylan and repressed by glucose at the transcriptional level. The promoter of this gene has a modular structure, with eight putative XlnR binding sites in tandem (XlnR module), and upstream from them, four putative CreA binding sites (CreA module). Promoter fragments containing different modules were inserted into a plasmid, pAN49-1, which contains a basal fungal promoter linked to a reporter gene (lacZ) and its expression was studied in vivo in Aspergillus nidulans. The XlnR module is able to trigger high β-galactosidase activity in the presence of xylan, but the lack of most XlnR sites notoriously reduces this enzymatic activity. No enzyme induction is observed if the orientation of the promoter fragment is inverted. The presence of the CreA module is necessary for glucose repression when β-galactosidase activity is previously induced by xylan. However, when transformant strains containing the XlnR module but lacking all CreA sites were grown in glucose without pre-induction in xylan, a low β-galactosidase activity was observed compared with the same transformants grown in xylan. These results agree with a double-lock regulatory mechanism for both direct and indirect repression of xylanolytic genes by glucose.

AB - In Penicillium purpurogenum, the gene encoding endoxylanase B (xynB) is highly expressed by xylan and repressed by glucose at the transcriptional level. The promoter of this gene has a modular structure, with eight putative XlnR binding sites in tandem (XlnR module), and upstream from them, four putative CreA binding sites (CreA module). Promoter fragments containing different modules were inserted into a plasmid, pAN49-1, which contains a basal fungal promoter linked to a reporter gene (lacZ) and its expression was studied in vivo in Aspergillus nidulans. The XlnR module is able to trigger high β-galactosidase activity in the presence of xylan, but the lack of most XlnR sites notoriously reduces this enzymatic activity. No enzyme induction is observed if the orientation of the promoter fragment is inverted. The presence of the CreA module is necessary for glucose repression when β-galactosidase activity is previously induced by xylan. However, when transformant strains containing the XlnR module but lacking all CreA sites were grown in glucose without pre-induction in xylan, a low β-galactosidase activity was observed compared with the same transformants grown in xylan. These results agree with a double-lock regulatory mechanism for both direct and indirect repression of xylanolytic genes by glucose.

KW - Aspergillus nidulans

KW - CreA sites

KW - In vivo expression

KW - Penicillium purpurogenum

KW - Promoter fragments

KW - XlnR sites

UR - http://www.scopus.com/inward/record.url?scp=50849088302&partnerID=8YFLogxK

U2 - 10.1007/s00294-008-0205-y

DO - 10.1007/s00294-008-0205-y

M3 - Article

C2 - 18661134

AN - SCOPUS:50849088302

VL - 54

SP - 133

EP - 141

JO - Current Genetics

JF - Current Genetics

SN - 0172-8083

IS - 3

ER -