Molecular cloning of rat leukemia inhibitory factor receptor alpha-chain gene and its expression during pregnancy.
Leukemia inhibitory factor (LIF) is a secreted glycoprotein and a pluripotent growth factor that acts on diverse cell systems. LIF transmits its effects via binding to transmembrane receptors, of which both high- and low-affinity forms have been identified. In this study, we analyzed the structure and expression of rat LIF receptor alpha-chain (rLIFR alpha) cDNA. A full-length clone of the cDNA encoding the membrane-bound form of rLIFR alpha protein was prepared by a combination of LA-PCR and 5' RACE using DNA reverse-transcribed from total RNA isolated from the livers of day-12 and day-14 pregnant rats as templates. The nucleotide sequence of a full-length clone was determined and further confirmed by analysis of shorter DNA fragment prepared by PCR using pfu polymerase. The gene for rLIFR alpha encodes a 1093 amino acid residue protein. The rLIFR alpha protein shows a high degree of similarity to mouse and human LIF receptor alpha-chain protein (89% and 76% amino acid sequence identities, respectively). Only one molecular species of mRNA for the rLIFR alpha gene was detected in the liver and placenta. rLIFR alpha was expressed in liver of both non-pregnant and pregnant rats. The level of mRNA for the rLIFR alpha gene in placenta was maximum on day 16 of pregnancy.
Aikawa
J
J
Laboratory of Synthetic Cellular Chemistry, RIKEN (the Institute of Physical and Chemical Research), Saitama, Japan. akemi@postman.riken.go.jp
Ikeda-Naiki
S
S
Ohgane
J
J
Min
K S
KS
Imamura
T
T
Sasai
K
K
Shiota
K
K
Ogawa
T
T
D000595:Amino Acid Sequence; D000818:Animals; D003001:Cloning, Molecular / Q000379:methods; D018076:DNA, Complementary / Q000235:genetics*; D004259:DNA-Directed DNA Polymerase; D005260:Female; D005786:Gene Expression Regulation / Q000502:physiology*; D005796:Genes / Q000235:genetics; D006131:Growth Inhibitors; D015850:Interleukin-6; D053605:Leukemia Inhibitory Factor; D053680:Leukemia Inhibitory Factor Receptor alpha Subunit; D008222:Lymphokines; D008969:Molecular Sequence Data; D009928:Organ Specificity; D016133:Polymerase Chain Reaction / Q000379:methods; D011247:Pregnancy; D012333:RNA, Messenger / Q000032:analysis; D051381:Rats; D018121:Receptors, Cytokine / Q000235:genetics*; D053606:Receptors, OSM-LIF; D017422:Sequence Analysis, DNA; D017386:Sequence Homology, Amino Acid
D018076:DNA, Complementary; D006131:Growth Inhibitors; D015850:Interleukin-6; C507837:LIF protein, human; C507840:LIFR protein, human; D053605:Leukemia Inhibitory Factor; D053680:Leukemia Inhibitory Factor Receptor alpha Subunit; C507838:Lif protein, mouse; C507841:Lifr protein, mouse; C507842:Lifr protein, rat; D008222:Lymphokines; D012333:RNA, Messenger; D018121:Receptors, Cytokine; D053606:Receptors, OSM-LIF; C072146:Pfu DNA polymerase; D004259:DNA-Directed DNA Polymerase