Functional characterization of Lilium lancifolium cold-responsive Zinc Finger Homeodomain (ZFHD) gene in abscisic acid and osmotic stress tolerance

Background. We have previously performed an analysis of the cold-responsive transcriptome in the mature leaves of tiger lily (Lilium lancifolium) by gene co-expression network identification. The results has revealed that a ZFHD gene, notated as encoding zinc finger homeodomain protein, may play an essential regulating role in tiger lily response to cold stress. Methods. A further investigation of the ZFHD gene (termed as LlZFHD4) responding to osmotic stresses, including cold, salt, water stresses, and abscisic acid (ABA) was performed in this study. Based on the transcriptome sequences, the coding region and 5′ promoter region of LlZFHD4 were cloned from mature tiger lily leaves. Stress response analysis was performed under continuous 4 °C, NaCl, PEG, and ABA treatments. Functional characterization of LlZFHD4 was conducted in transgenic Arabidopsis, tobacco, and yeast. Results. LlZFHD4 encodes a nuclear-localized protein consisting of 180 amino acids. The N-terminal region of LlZFHD4 has transcriptional activation activity in yeast. The 4 °C, NaCl, PEG, and ABA treatments induced the expression of LlZFHD4. Several stress- or hormone-responsive cis-acting regulatory elements (T-Box, BoxI. and ARF) and binding sites of transcription factors (MYC, DRE and W-box) were found in the core promoter region (789 bp) of LlZFHD4. Also, the GUS gene driven by LlZFHD4 promoter was up-regulated by cold, NaCl, water stresses, and ABA in Arabidopsis. Overexpression of LlZFHD4 improved cold and drought tolerance in transgenic Arabidopsis; higher survival rate and better osmotic adjustment capacity were observed in LlZFHD4 transgenic plants compared to wild type (WT) plants under 4 °C and PEG conditions. However, LlZFHD4 transgenic plants were less tolerant to salinity and more hypersensitive to ABA compared to WT plants. The transcript levels of stress- and ABA-responsive genes were much more up-regulated in LlZFHD4 transgenic Arabidopsis than WT. These results indicate LlZFHD4 is involved in ABA signaling pathway and plays a crucial role in regulating the response of tiger lily to cold, salt and water stresses.

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Characterization of Hepatic-specific Regulatory Elements in the Promoter Region of the Human Cholesterol 7α-Hydroxylase Gene

Journal of Biological Chemistry ◽

10.1074/jbc.272.6.3444 ◽

1997 ◽

Vol 272 (6) ◽

pp. 3444-3452 ◽

Cited By ~ 41

Author(s):

Allen D. Cooper ◽

Jean Chen ◽

Mary Jane Botelho-Yetkinler ◽

Yicheng Cao ◽

Takahiro Taniguchi ◽

...

Keyword(s):

Promoter Region ◽

Regulatory Elements ◽

Hydroxylase Gene

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Abstract 050: Fine-mapping and Bioinformatics Characterization of QT Interval Loci in African Americans: The Population Architecture Using Genomics and Epidemiology (PAGE) Study

Circulation ◽

10.1161/circ.125.suppl_10.a050 ◽

2012 ◽

Vol 125 (suppl_10) ◽

Author(s):

Christy L Avery ◽

Praveen Sethupathy ◽

Steven Buyske ◽

Q. C He ◽

Dan Y Lin ◽

...

Keyword(s):

African American ◽

African Americans ◽

Fine Mapping ◽

Qt Interval ◽

Functional Characterization ◽

Regulatory Elements ◽

European Ancestry ◽

Functional Evaluation ◽

Human Cardiomyocytes

The QT interval (QT) is a heritable trait and its prolongation is an established risk factor for ventricular tachyarrhythmia and sudden cardiac death. Most genetic studies of QT have examined populations of European ancestry, although the increased genetic diversity in populations of African descent provides opportunity for fine-mapping, which can help narrow association signals and identify candidates for functional characterization. We examined whether eleven previously identified QT loci comprising 6,681 variants on the Illumina Metabochip array were associated with QT in 7,516 African American participants from the Atherosclerosis Risk in Communities study and Women’s Health Initiative clinical trial. Among associated loci, we used conditional analyses and queried bioinformatics databases to identify and functionally categorize signals. We identified nine of the eleven QT loci in African American populations ( P <0.0045 under an additive genetic model adjusting for ancestry and demographic characteristics: NOS1AP, ATP1B1, SCN5A, SLC35F1, KCNH2, KCNQ1, LITAF, NDRG4, and RFFL ). We also identified two independent secondary signals in NOS1AP and ATP1B1 ( P < 7.4x10 −6 ). Conditional analyses adjusting for published loci in European populations demonstrated that eight of these eleven SNPs (nine primary; two secondary) were independent of previously reported SNPs. We then performed the first bioinformatics-based functional characterization of QT loci using the eleven primary and secondary variants and SNPs in strong LD (r 2 > 0.5) among these African American participants. Only the SCN5A locus included a non-synonymous coding variant (rs1805124, H558R, r 2 = 0.7 with primary SNP rs9871385, P = 4.7x10 −4 ). The remaining ten loci harbored variants located exclusively within non-coding regions. Specifically, three contained SNPs within candidate long-range regulatory elements in human cardiomyocytes, five were in or near annotated promoter regions, and the remaining two were in un-annotated, but highly conserved non-coding elements. Several of the QT risk alleles at these SNPs significantly alter the predicted binding affinity for transcription factors, such as TBX5 and AhR, which have been previously implicated in cardiac formation and function. In summary, the findings provide compelling evidence that the same genes influence variation in QT across global populations and that additional, independent signals exist in African Americans. Moreover, of those SNPs identified as strong candidates for functional evaluation, the majority implicate gene regulatory dysfunction in QT prolongation.

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Functional Characterization of Variants inMC4RGene Promoter Region Found in Obese Children

The Journal of Clinical Endocrinology & Metabolism ◽

10.1210/jc.2013-3711 ◽

2014 ◽

Vol 99 (5) ◽

pp. E931-E935 ◽

Cited By ~ 2

Author(s):

Karen Mei Ling Tan ◽

Shu Qin Delicia Ooi ◽

Siong Gim Ong ◽

Charmaine Shuyi Kwan ◽

Raymond Ming En Chan ◽

...

Keyword(s):

Promoter Region ◽

Functional Characterization ◽

Obese Children

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Structural and functional characterization of the promoter region of the mouse c-Ki-ras gene

Molecular and Cellular Biology ◽

10.1128/mcb.7.7.2592-2596.1987 ◽

1987 ◽

Vol 7 (7) ◽

pp. 2592-2596

Author(s):

E K Hoffman ◽

S P Trusko ◽

N Freeman ◽

D L George

Keyword(s):

Promoter Region ◽

Promoter Activity ◽

Functional Characterization ◽

Genetic Material ◽

Bidirectional Promoter ◽

Adrenocortical Tumor ◽

Ras Gene ◽

Untranslated Exon ◽

Size Heterogeneity

We isolated and characterized the 5' region of the mouse c-Ki-ras gene, including a 5' untranslated exon (exon 0). These studies used genetic material from Y1 mouse adrenocortical tumor cells in which the c-Ki-ras gene is amplified and overexpressed. Our data demonstrate that transcription initiates at multiple sites, predicting size heterogeneity at the 5' ends of the c-Ki-ras mRNAs. Using transient expression assays, we identified a genomic fragment within the 5' region which exhibits bidirectional promoter activity.

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