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1,25-dihydroxyvitamin D3 regulation ...

Watana, Surawut.

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1,25-dihydroxyvitamin D3 regulation of osteocalcin mRNA stability: A search for cis- and trans-acting determinants.

紀錄類型:	書目-電子資源 : Monograph/item
正題名/作者:	1,25-dihydroxyvitamin D3 regulation of osteocalcin mRNA stability: A search for cis- and trans-acting determinants./
作者:	Watana, Surawut.
面頁冊數:	203 p.
附註:	Source: Dissertation Abstracts International, Volume: 64-05, Section: B, page: 2143.
Contained By:	Dissertation Abstracts International64-05B.
標題:	Health Sciences, Pharmacology. -
電子資源:	http://pqdd.sinica.edu.tw/twdaoapp/servlet/advanced?query=3089627

1,25-dihydroxyvitamin D3 regulation of osteocalcin mRNA stability: A search for cis- and trans-acting determinants.
Watana, Surawut.

1,25-dihydroxyvitamin D3 regulation of osteocalcin mRNA stability: A search for cis- and trans-acting determinants. - 203 p.

Source: Dissertation Abstracts International, Volume: 64-05, Section: B, page: 2143.

Thesis (Ph.D.)--The University of Wisconsin - Madison, 2003.

<italic>Cis</italic>- and <italic>trans</italic>-acting determinants for the 1,25(OH)2D3 regulation of osteocalcin mRNA stability were investigated. Deletion mutation analyses were performed with constitutively expressed osteocalcin mRNA in U937 cells. Deletion of sequences in the 5<super> ′</super>-untranslated region (5<super>′</super>-UTR) resulted in stable osteocalcin mRNA both in the presence and absence of 1,25(OH) 2D3. The results suggest the existence of a destabilizing/stabilizing element conferring the stability of osteocalcin mRNA in the 5<super>′ </super>-UTR. Further studies utilizing ribonuclease probing and MFOLD prediction suggest that a sequence consisting of the 5<super>′</super>-UTR and the first 30 nucleotides in the osteocalcin coding region forms a stem-loop structure. To further determine a role of the osteocalcin 5<super>′ </super>-UTR stem-loop, U937 cells were transfected with a chimeric gene containing sequences from the stem-loop and the <italic>E. coli</italic> choramphenicol acetyltransferase (CAT) gene, followed by examining the effect of 1,25(OH) 2D3 on the decay rate of the chimeric mRNA. The stability of the mRNA transcribed from the chimeric construct was shown to be regulated by 1,25(OH)2D3. In addition, RNA electrophoretic mobility shift analyses (R-EMSAs) show multiprotein complexes that specifically interact with the osteocalcin 5<super>′</super>-UTR stern-loop. Ultraviolet (UV) cross-liking analyses of the stem-loop reveal ribonucleoprotein complexes forming with ROS 17/2.8 cellular lysates ranging from 50 to 130 kDa, with the major complex migrating at &sim;50 kDa. These results suggest that multiprotein complexes may play a role in the 1,25(OH)2D3 regulation of ostelcalcin mRNA stability. To further elucidate the significance of the primary and secondary structure of the stem-loop in the regulation of osteocalcin mRNA stability, several mutations were made in the stem-loop. The results suggest that the single-stranded region of the stem-loop may be responsible for the binding of RNA-binding proteins as well as the 1,25(OH)2D 3—mediated stability of osteocalcin mRNA. Moreover, the pyrimidine-rich region corresponding to nucleotides +8 to +26 in the stem-loop also may play a role in RNA-protein recognition, thereby mediating an effect of 1,25(OH) 2D3 in enhancing the stability of osteocalcin mRNA.Subjects--Topical Terms:

1017717
Health Sciences, Pharmacology.

1,25-dihydroxyvitamin D3 regulation of osteocalcin mRNA stability: A search for cis- and trans-acting determinants.
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