Identification of regulatory elements in the AGT1 promoter of ale and lager strains of brewer's yeast

Virve Vidgren (Corresponding Author), Matti Kankainen, John Londesborough, Laura Ruohonen

Research output: Contribution to journalArticleScientificpeer-review

15 Citations (Scopus)

Abstract

Agt1 is an interesting α‐glucoside transporter for the brewing industry, as it efficiently transports maltotriose, a sugar often remaining partly unused during beer fermentation. It has been shown that on maltose the expression level of AGT1 is much higher in ale strains than in lager strains, and that glucose represses the expression, particularly in the ale strains. In the present study the regulatory elements of the AGT1 promoter of one ale and two lager strains were identified by computational methods. Promoter regions up to 1.9 kbp upstream of the AGT1 gene were sequenced from the three brewer's yeast strains and the laboratory yeast strain CEN.PK‐1D. The promoter sequence of the laboratory strain was identical to the AGT1 promoter of strain S288c of the Saccharomyces Genome Database, whereas the promoter sequences of the industrial strains diverged markedly from the S288c strain. The AGT1 promoter regions of the ale and lager strains were for the most part identical to each other, except for one 22 bp deletion and two 94 and 95 bp insertions in the ale strain. Computational analyses of promoter elements revealed that the promoter sequences contained several Mig1‐ and MAL‐activator binding sites, as was expected. However, some of the Mig1 and MAL‐activator binding sites were located on the two insertions of the ale strain, and thus offered a plausible explanation for the different expression pattern of the AGT1 gene in the ale strains. Accordingly, functional analysis of A60 ale and A15 lager strain AGT1 promoters fused to GFP (encoding the green fluorescent protein) showed a significant difference in the ability of these two promoters to drive GFP expression. Under the control of the AGT1 promoter of the ale strain the emergence of GFP was strongly induced by maltose, whereas only a low level of GFP was detected with the construct carrying the AGT1 promoter of the lager strain. Thus, the extra MAL‐activator binding element, present in the AGT1 promoter of the ale strain, appears to be necessary to reach a high level of induction by maltose. Both AGT1 promoters were repressed by glucose but their derepression was different, possibly due to a distinct distribution of Mig1 elements in these two promoters.
Original languageEnglish
Pages (from-to)579-594
Number of pages16
JournalYeast
Volume28
Issue number8
DOIs
Publication statusPublished - 2011
MoE publication typeA1 Journal article-refereed

Fingerprint

Beer
Green Fluorescent Proteins
Yeast
Maltose
Saccharomyces cerevisiae
Genetic Promoter Regions
Binding Sites
Glucose
Saccharomyces
Glucosides
Genes
Fermentation
Industry
Yeasts
Proteins
Genome
Databases
Binding sites
Brewing
Functional analysis

Keywords

  • AGT1 promoter
  • brewer's yeast strains
  • MAL-activator
  • Mig1

Cite this

Vidgren, Virve ; Kankainen, Matti ; Londesborough, John ; Ruohonen, Laura. / Identification of regulatory elements in the AGT1 promoter of ale and lager strains of brewer's yeast. In: Yeast. 2011 ; Vol. 28, No. 8. pp. 579-594.
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Identification of regulatory elements in the AGT1 promoter of ale and lager strains of brewer's yeast. / Vidgren, Virve (Corresponding Author); Kankainen, Matti; Londesborough, John; Ruohonen, Laura.

In: Yeast, Vol. 28, No. 8, 2011, p. 579-594.

Research output: Contribution to journalArticleScientificpeer-review

TY - JOUR

T1 - Identification of regulatory elements in the AGT1 promoter of ale and lager strains of brewer's yeast

AU - Vidgren, Virve

AU - Kankainen, Matti

AU - Londesborough, John

AU - Ruohonen, Laura

PY - 2011

Y1 - 2011

N2 - Agt1 is an interesting α‐glucoside transporter for the brewing industry, as it efficiently transports maltotriose, a sugar often remaining partly unused during beer fermentation. It has been shown that on maltose the expression level of AGT1 is much higher in ale strains than in lager strains, and that glucose represses the expression, particularly in the ale strains. In the present study the regulatory elements of the AGT1 promoter of one ale and two lager strains were identified by computational methods. Promoter regions up to 1.9 kbp upstream of the AGT1 gene were sequenced from the three brewer's yeast strains and the laboratory yeast strain CEN.PK‐1D. The promoter sequence of the laboratory strain was identical to the AGT1 promoter of strain S288c of the Saccharomyces Genome Database, whereas the promoter sequences of the industrial strains diverged markedly from the S288c strain. The AGT1 promoter regions of the ale and lager strains were for the most part identical to each other, except for one 22 bp deletion and two 94 and 95 bp insertions in the ale strain. Computational analyses of promoter elements revealed that the promoter sequences contained several Mig1‐ and MAL‐activator binding sites, as was expected. However, some of the Mig1 and MAL‐activator binding sites were located on the two insertions of the ale strain, and thus offered a plausible explanation for the different expression pattern of the AGT1 gene in the ale strains. Accordingly, functional analysis of A60 ale and A15 lager strain AGT1 promoters fused to GFP (encoding the green fluorescent protein) showed a significant difference in the ability of these two promoters to drive GFP expression. Under the control of the AGT1 promoter of the ale strain the emergence of GFP was strongly induced by maltose, whereas only a low level of GFP was detected with the construct carrying the AGT1 promoter of the lager strain. Thus, the extra MAL‐activator binding element, present in the AGT1 promoter of the ale strain, appears to be necessary to reach a high level of induction by maltose. Both AGT1 promoters were repressed by glucose but their derepression was different, possibly due to a distinct distribution of Mig1 elements in these two promoters.

AB - Agt1 is an interesting α‐glucoside transporter for the brewing industry, as it efficiently transports maltotriose, a sugar often remaining partly unused during beer fermentation. It has been shown that on maltose the expression level of AGT1 is much higher in ale strains than in lager strains, and that glucose represses the expression, particularly in the ale strains. In the present study the regulatory elements of the AGT1 promoter of one ale and two lager strains were identified by computational methods. Promoter regions up to 1.9 kbp upstream of the AGT1 gene were sequenced from the three brewer's yeast strains and the laboratory yeast strain CEN.PK‐1D. The promoter sequence of the laboratory strain was identical to the AGT1 promoter of strain S288c of the Saccharomyces Genome Database, whereas the promoter sequences of the industrial strains diverged markedly from the S288c strain. The AGT1 promoter regions of the ale and lager strains were for the most part identical to each other, except for one 22 bp deletion and two 94 and 95 bp insertions in the ale strain. Computational analyses of promoter elements revealed that the promoter sequences contained several Mig1‐ and MAL‐activator binding sites, as was expected. However, some of the Mig1 and MAL‐activator binding sites were located on the two insertions of the ale strain, and thus offered a plausible explanation for the different expression pattern of the AGT1 gene in the ale strains. Accordingly, functional analysis of A60 ale and A15 lager strain AGT1 promoters fused to GFP (encoding the green fluorescent protein) showed a significant difference in the ability of these two promoters to drive GFP expression. Under the control of the AGT1 promoter of the ale strain the emergence of GFP was strongly induced by maltose, whereas only a low level of GFP was detected with the construct carrying the AGT1 promoter of the lager strain. Thus, the extra MAL‐activator binding element, present in the AGT1 promoter of the ale strain, appears to be necessary to reach a high level of induction by maltose. Both AGT1 promoters were repressed by glucose but their derepression was different, possibly due to a distinct distribution of Mig1 elements in these two promoters.

KW - AGT1 promoter

KW - brewer's yeast strains

KW - MAL-activator

KW - Mig1

U2 - 10.1002/yea.1888

DO - 10.1002/yea.1888

M3 - Article

VL - 28

SP - 579

EP - 594

JO - Yeast

JF - Yeast

SN - 0749-503X

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ER -