Cloning and functional characterization of the 5′ regulatory region of ovine Hormone Sensitive Lipase (HSL) gene

Lampidonis, Antonis D.; Stravopodis, Dimitrios J.; Voutsinas, Gerassimos E.; Messini-Nikolaki, Niki; Stefos, Georgios C; Margaritis, Lukas H.; Argyrokastritis, Alexandros; Bizelis, Iosif; Rogdakis, E.

doi:10.1016/j.gene.2008.09.001

Cited by 20 publications

(14 citation statements)

References 52 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…Significant effects determined by two-way analysis of variance indicated by *P \ 0.10 or NS not significant Despite the absence of differences between treatments for whole-body cortisol concentrations, which were in accordance with values reported by Szisch et al [1] for seabream, GR mRNA levels were higher in SBL diet fed fish. A similar trend was noted for HSL which has been identified as a glucocorticoid-sensitive gene in mammals [38], containing glucocorticoid responsive elements (GREs) in its promoter [39,40]. This is also likely in fish as enhanced lipolytic activity and increased free fatty acid levels in the plasma are generally associated to the stress response (reviewed by Mommsen et al [12]).…”

Section: Discussionmentioning

confidence: 58%

Dietary Lecithin Source Affects Growth Potential and Gene Expression in Sparus aurata Larvae

Martins

Estévez

Stickland

et al. 2010

Lipids

View full text Add to dashboard Cite

Soybean lecithin (SBL), used as a phospholipid source in larval fish diets, may compromise growth and survival in marine species, and affect gene expression, due to differences in fatty acid composition relative to marine lecithins (ML). The potential of SBL as a phospholipid source in gilthead seabream microdiets as compared to ML was evaluated. Two stocking densities were tested in order to exacerbate possible dietary effects: 5 and 20 larvae L(-1). Larvae reflected dietary fatty acid profiles: linoleic acid was higher, whereas eicosapentaenoic and arachidonic acids were lower in SBL fed groups than in ML fed larvae. Highest stocking density decreased survival, and led to elevated saturates and lower docosahexaenoic acid levels in polar lipid. Muscle histology observations showed hindered growth potential in SBL fed larvae. Despite similar cortisol levels between treatments, higher glucocorticoid receptor (GR), as well as hormone-sensitive lipase (HSL) mRNA levels in SBL fed groups revealed a role for fatty acids in gene regulation. Further analysed genes suggested these effects were independent from the hypothalamus-pituitary-interrenal axis control and the endocannabinoid system. Cyclooxygenase-2 and gluconeogenesis seemed unaffected. For the first time in fish, a link between dietary lecithin nature and HSL gene transcription, perhaps regulated through GR fatty acid-induced activation, is suggested. Enhanced lipolytic activity could partly explain lower growth in marine fish larvae when dietary ML is not provided.

show abstract

Section: Discussionmentioning

confidence: 58%

Dietary Lecithin Source Affects Growth Potential and Gene Expression in Sparus aurata Larvae

Martins

Estévez

Stickland

et al. 2010

Lipids

View full text Add to dashboard Cite

show abstract

“…A TATAAA sequence, as a classical TATA box, locates at -30 bp upstream of the transcription starting point, which was in agreement with the consensus distance generally found in promoter regions. In some cases, other consensus sequences, such as TTATTT, could act as substitutes for the classical TATA box (Reed and Gibson 1994;Lampidonis et al 2008). In eukaryotes, a functional CAAT box is typically found about -75 bp upstream the transcriptional starting site.…”

Section: Resultsmentioning

confidence: 99%

Cloning and sequence analysis of a novel xylanase gene, Auxyn10A, from Aspergillus usamii

Wang

Zhang

et al. 2011

Biotechnol Lett

View full text Add to dashboard Cite

A full-length cDNA sequence, encoding a novel endo-1,4-β-D: -xylanase (AuXyn10A) of Aspergillus usamii, was obtained by using rapid amplification of cDNA ends (RACE) methods and cloned into the pUCm-T vector, followed by DNA sequencing. The cDNA gene, designated as Auxyn10A, is 1,235 bp in length harboring 5'- and 3'-non-encoding regions, as well as an ORF of 984 bp that encodes a 19-aa signal peptide, a 6-aa propeptide and a 302-aa mature peptide with a calculated MW of 32,756 Da. The AuXyn10A displays high similarity to the xylanases of Aspergillus niger, Aspergillus kawachii and Aspergillus niger, members of the glycoside hydrolase family 10. Its three-dimensional structure was predicted using http://swiss-model.expasy.org/on-line programs based on the crystal structure of Penicillium simplicissimum xylanase (1B30_A) from the family 10. The complete DNA gene was cloned from the genomic DNA of A. usamii using conventional PCR and hairpin structure-mediated PCR techniques. The DNA gene is 2,255 bp in length, containing a 510 bp of 5'-flanking promoter region and a 1,745 bp of downstream fragment that consists of ten exons and nine short introns ranging from 52 to 62 bp.

show abstract

“…A TATTTAA sequence in the gene Aucel12A that functions as a substitute for the classical TATA box (TATAAA) which is located -25 bp upstream the transcription starting point, which was in agreement with the consensus distance generally found in eukaryote promoters. Other consensus sequences such as TTATTT could also act as substitutes for the classical TATA box [11,16]. In eukaryotes, the functional CAAT box is typically found about -75 bp upstream from the transcription starting point.…”

Section: Dna Sequence Characterization Of the Gene Aucel12amentioning

confidence: 99%

Cloning and bioinformatics analysis of an endoglucanase gene (Aucel12A) from Aspergillus usamii and its functional expression in Pichia pastoris

Shi

Yin

et al. 2012

Journal of Industrial Microbiology and Biotechnology

View full text Add to dashboard Cite

Using 3' and 5' rapid amplification of cDNA ends methods, the full-length cDNA sequence encoding an endo-1,4-β-glucanase of Aspergillus usamii E001 (abbreviated as AuCel12A) was amplified from the total RNA. The clone cDNA sequence of the gene encoding the AuCel12A, named as Aucel12A, is 1,027 bp in length harboring 5' and 3' non-coding regions, as well as a 720 bp of open reading frame that encodes a 16-aa signal peptide, and a 223-aa mature AuCel12A with a theoretical M.W. of 24,294 Da, a calculated pI of 4.15, and one putative N-glycosylation site. The complete DNA sequence of the gene Aucel12A was amplified from the genomic DNA of A. usamii E001 by using the conventional PCR and pUCm-T vector-mediated PCR initially developed in our lab. The clone DNA sequence is 1,576 bp in length, consisting of a 5' flanking regulatory region, three exons, and two introns with sizes of 50 and 66 bp. The cDNA fragment encoding the mature AuCel12A was expressed in a fully active form in Pichia pastoris. One P. pastoris transformant expressing the highest recombinant AuCel12A (rAuCel12A) activity, labeled as P. pastoris GSCel2-1, was chosen for subsequent studies. Integration of the Aucel12A into P. pastoris genome was confirmed by PCR analysis using 5'- and 3'-AOX1 primers. SDS-PAGE and enzyme activity assays demonstrated that the rAuCel12A, a glycosylated protein with an apparent M.W. of 27.0 kDa and a carbohydrate content of 4.82%, was secreted into the culture medium. The purified rAuCel12A displayed the highest activity at pH 5.0 and 60°C. It was highly stable at a pH range of 3.5-7.0, and at a temperature of 55°C or below. Its activity was not significantly affected by an array of metal ions and EDTA, but inhibited by Ag⁺, Hg²⁺ and Fe²⁺. The K(m) and V(max) of the rAuCel12A, towards carboxymethylcellulose-Na (CMC-Na) at pH 5.0 and 50°C were 4.85 mg/ml and 160.5 U/mg, respectively.

show abstract

Cloning and functional characterization of the 5′ regulatory region of ovine Hormone Sensitive Lipase (HSL) gene

Cited by 20 publications

References 52 publications

Dietary Lecithin Source Affects Growth Potential and Gene Expression in Sparus aurata Larvae

Dietary Lecithin Source Affects Growth Potential and Gene Expression in Sparus aurata Larvae

Cloning and sequence analysis of a novel xylanase gene, Auxyn10A, from Aspergillus usamii

Cloning and bioinformatics analysis of an endoglucanase gene (Aucel12A) from Aspergillus usamii and its functional expression in Pichia pastoris

Contact Info

Product

Resources

About