Molecular cloning and characterization of an amphibian progesterone receptor from Rana dybowskii.
Progesterone plays a pivotal role in the regulation of reproduction in all vertebrates and binds to nuclear hormone receptor, one of ligand-dependent transcription factors. Although avian and mammalian progesterone receptors (PR) have been well characterized, detail structure and function of amphibian progesterone receptor in wild frog is poorly studied yet. Here we report the cloning and characterization of a novel progesterone receptor from the Korean frog, Rana dybowskii. The R. dybowskii progesterone receptor (dyPR, GenBank Accession No. AF431813) cDNA isolated from testis encodes a protein of 711 amino acids which shows approximately 60% overall identity with the Xenopus progesterone receptor. Reverse transcriptase-polymerase chain reaction (RT-PCR) demonstrates that dyPR is expressed in all the tissues examined. Electrophoretic mobility shift assays demonstrate that this receptor specifically binds to a progesterone response element (PRE), and transient transfection studies demonstrate that dyPR significantly activates the transcription of a PRE containing reporter element. Finally, confocal microscopy demonstrates the localization of this protein in nucleus, cytoplasm, and plasma membrane in transiently transfected CV-1 cell. These results indicate that dyPR cDNA encodes a classical progesterone receptor and molecular characterization of dyPR may provide us new information about the evolution of steroid hormone receptor.
Wang
Li
L
Hormone Research Center, Chonnam National University, Kwangju 500-757, Republic of Korea.
Sanyal
Sabyasachi
S
Oh
Da Young
DY
Oh
Da Young
da Y
Kim
Joon-Young
JY
Ju
Jung Won
JW
Song
Kwang-Hoon
KH
Kim
Jung Woo
JW
Kwon
Hyuk Bang
HB
Choi
Hueng-Sik
HS
D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D003001:Cloning, Molecular; D018076:DNA, Complementary / Q000032:analysis; D004268:DNA-Binding Proteins / Q000378:metabolism; D008969:Molecular Sequence Data; D011898:Ranidae; D011980:Receptors, Progesterone / Q000235:genetics* / Q000378:metabolism; D020133:Reverse Transcriptase Polymerase Chain Reaction; D017386:Sequence Homology, Amino Acid; D013347:Subcellular Fractions; D015533:Transcriptional Activation
D018076:DNA, Complementary; D004268:DNA-Binding Proteins; D011980:Receptors, Progesterone