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Translational and structural analysis of the shortest legume ENOD40 gene in Lupinus luteus

100%

Podkowinski J., Zmienko A., Florek B., Wojciechowski P., Rybarczyk A., Wrzesinski J., Ciesiolka J., Blazewicz J., Kondorosi A., Crespi M., Legocki A.

Acta Biochimica Polonica

2009

tom 56

nr 1

89-102

Two early nodulin 40 (enod40) genes, ENOD40-1, the shortest legume ENOD40 gene, and ENOD40-2, were isolated from Lupinus luteus, a legume with indeterminate nodules. Both genes were expressed at similar levels during symbiosis with nitrogen-fixing bacteria. ENOD40 phylogeny clustered the L. luteus genes with legumes forming determinate nodules and revealed peptide similarities. The ENOD40-1 small ORF A fused to a reporter gene was efficiently expressed in plant cells, indicating that the start codon is recognized for translation. The ENOD40-1 RNA structure predicted based on Pb(II)-induced cleavage and modeling revealed four structurally conserved domains, an absence of domain 4 characteristic for legumes of indeterminate nodules, and interactions between the conserved region I and a region located upstream of domain 6. Domain 2 contains Mg(II) ion binding sites essential for organizing RNA secondary structure. The differences between L. luteus and Glycine max ENOD40 RNA models suggest the possibility of a switch between two structural states of ENOD40 transcript.

The structural and functional analysis of the human HSPA2 gene promoter region

80%

Piglowski W., Nowak R., Krawczyk Z., Scieglinska D.

Acta Biochimica Polonica

2007

tom 54

nr 1

HSPA2 is a human counterpart of the testis-specific rodent Hst70/Hsp70.2 gene. In contrast to the latter, the expression of the human HSPA2 gene is not limited to the testis, and recent data show that human tumor cells can express this gene at significant levels. The characteristics of HSPA2 expression suggests that it can influence the phenotype and survival of cancer cells similarly as overexpression of major members of the HSP70 gene family. Until now, neither the structure of the transcription unit of the human HSPA2 gene has been established nor a functional analysis of its promoter performed. In this study we established that the human HSPA2 gene, in contrast to its rodent counterparts, is intronless and has a single transcription start site. We also show that the same type of HSPA2 transcripts are synthesized in the testes and in cancer cell lines. In order to perform a functional study of the HSPA2 promoter, we used a transient transfection assay and found that the 392 bp fragment upstream of the ATG codon was a minimal region required for efficient transcription, while a 150 bp deletion from the 5' end of this region dramatically reduced the promoter activity. Delineation of the minimal promoter is a basic step toward identifiying the cis and trans elements involved in the regulation of the HSPA2 gene expression in cancer cells.

Ograniczanie wyników

2 Acta Biochimica Polonica

1 Blazewicz J.

1 Ciesiolka J.

1 Crespi M.

1 Florek B.

1 Kondorosi A.

1 Krawczyk Z.

1 Legocki A.

1 Nowak R.

1 Piglowski W.

1 Podkowinski J.

1 Rybarczyk A.

1 Scieglinska D.

1 Wojciechowski P.

1 Wrzesinski J.

1 Zmienko A.

1 2009

1 2007

Wyniki wyszukiwania

Translational and structural analysis of the shortest legume ENOD40 gene in Lupinus luteus

The structural and functional analysis of the human HSPA2 gene promoter region