Nucleotide sequence of mouse IL-2 receptor cDNA and its comparison with the human IL-2 receptor sequence

Abstract

We have cloned cDNA encoding the mouse interleukin-2(IL-2) receptor from a murine T cell line, CTLL using human IL-2 receptor cDNA as probe. COS 7 cells transfected with the cDNA expressed the antigen recognized by the monoclonal antibody against the murine IL-2 receptor. The cDNA identified 4 species of mRNA (4.5, 3.5, 2.2 and 1.5 kb) of the mouse IL-2 receptor in CTLL cells. Difference in the length of mRNA seems to be ascribed to the variable length of the 3' untranslated sequence. Total nucleotide sequence ('1400 bp) of this cDNA was determined and compared with the human receptor. The nucleotide and amino acid sequences of the IL-2 receptor are 70% and 60%, respectively, homologous in average between the two species. The comparison has revealed several conserved regions localized to particular exons such as transmembrane and cytoplasmic portions, suggesting that these regions are important for receptor function and its regulation.

Overexpression of AML1 renders a T hybridoma resistant to T cell receptor-mediated apoptosis

“…Transfer of RNA onto a Hybond N membrane (Amersham) and the procedures of hybridization and washing were as described previously . The DNA fragments used to make probes were as follows; an EcoRI/AccI fragment from murine Fas cDNA (nt 1 ± 1250, Watanabe-Fukunaga et al, 1992b), a 940 bp XbaI fragment from murine Fas-ligand cDNA harboring the open reading frame , a 430 bp PstI fragment from murine IL-2Ra cDNA harboring the amino-terminal half portion of the open reading frame (Shimizu et al, 1985) and a KpnI fragment of b-actin cDNA (nt 1 ± 1150, Minty et al, 1981). The DNA fragments to make probes for detecting the Bcl-2, Bcl-x and Bax transcripts were prepared by PCR ampli®cation using the P19 cell or mouse brain derived cDNAs as templates.…”

Molecular cloning and expression of chicken C-terminal Src kinase: lack of stable association with c-Src protein.

“…Even under the mildest conditions (lx SSC/0.1% SDS; 50°C), only two additional very faint bands were detected with the csk probe. Under these hybridization conditions, a gene with a homology of =69% can be detected (29).…”

Interleukin-2 Carbohydrate Recognition Modulates CTLL-2 Cell Proliferation

“…Since Endo H hydrolyzes high-mannose type glycans including Man 4 -9 GlcNAc 2 and hybrid-type glycans (23), whereas Endo D hydrolyzes Man 3-5 GlcNAc 2 (15,24) and since IL-2-dependent proliferation of CTLL-2 cells was inhibited by the addition of Man 5 GlcNAc 2 Asn or Man 6 GlcNAc 2 Asn, the carbohydrate structure of IL-2R␣ to which IL-2 binds appears to include Man 5 GlcNAc 2 . These results suggest that only the IL-2R␣ subunit has the high-mannose type glycan with Man 5 GlcNAc 2 , among the components of the IL-2R complex in CTLL-2 cells, and that IL-2 bifunctionally binds a high-mannose type glycan and a specific peptide sequence of IL-2R␣, although all of the subunits of IL-2R have several potential N-glycosylation sites (25)(26)(27).…”

“…Since the nonglycosylated rhIL-2R␣ subunit recognizes Lys 35 , Lys 38 , Thr 42 , and Lys 43 residues in IL-2 (28), another peptide sequence of IL-2 may bind a high-mannose type glycan with Man 5 GlcNAc 2 which is linked to Asn 33 , Asn 43 , or Asn 200 of mouse IL-2R␣ (25). As soon as IL-2 bifunctionally binds to the IL-2R␣ subunit, formation of the IL-2⅐IL-2R␣ complex might occur resulting in a change in conformation of IL-2 which increases the accessibility to the IL-2R␤ and IL-2R␥ subunits.…”