Subtype-specific regulation of equilibrative nucleoside transporters by protein kinase CK2
| Autor: | James R. Hammond, Greg Vilk, Elizabeth A. Cooper, David W. Litchfield, Meaghan Stolk |
|---|---|
| Rok vydání: | 2005 |
| Předmět: |
2-Chloroadenosine
Bone Neoplasms Equilibrative nucleoside transporter 2 Nucleoside transporter Transfection Tritium Equilibrative nucleoside transporter 1 Polymerase Chain Reaction Biochemistry Substrate Specificity Equilibrative Nucleoside Transporter 1 Computer Systems Thioinosine Catalytic Domain Cell Line Tumor medicine Humans Equilibrative-Nucleoside Transporter 2 Casein Kinase II Protein kinase A Molecular Biology Osteosarcoma biology Nucleosides Equilibrative nucleoside transporter Cell Biology Adenosine Molecular biology Transport protein Gene Expression Regulation Neoplastic biology.protein Formycins Nucleoside Research Article medicine.drug |
| Zdroj: | Biochemical Journal. 386:281-289 |
| ISSN: | 1470-8728 0264-6021 |
| Popis: | Two subtypes of equilibrative transporters, es (equilibrative inhibitor-sensitive) and ei (equilibrative inhibitor-insensitive), are responsible for the majority of nucleoside flux across mammalian cell membranes. Sequence analyses of the representative genes, ENT1 {equilibrative nucleoside transporter 1; also known as SLC29A1 [solute carrier family 29 (nucleoside transporters), member 1]} and ENT2 (SLC29A2), suggest that protein kinase CK2-mediated phosphorylation may be involved in the regulation of es- and ei-mediated nucleoside transport. We used human osteosarcoma cells transfected with catalytically active or inactive alpha' and alpha subunits of CK2 to assess the effects of CK2 manipulation on nucleoside transport activity. Expression of inactive CK2alpha' (decreased CK2alpha' activity) increased the number of binding sites (approximately 1.5-fold) for the es-specific probe [3H]NBMPR ([3H]nitrobenzylthioinosine), and increased (approximately 1.8-fold) the V(max) for 2-chloro[3H]adenosine of the NBMPR-sensitive (es) nucleoside transporter. There was a concomitant decrease in the V(max) of the NBMPR-resistant (ei-mediated) uptake of 2-chloro[3H]adenosine. This inhibition of CK2alpha' activity had no effect, however, on either the K(D) of [3H]NBMPR binding or the K(m) of 2-chloro[3H]adenosine uptake. Quantitative PCR showed a transient decrease in the expression of both hENT1 (human ENT1) and hENT2 mRNAs within 4-12 h of induction of the inactive CK2alpha' subunit, but both transcripts had returned to control levels by 24 h. These data suggest that inhibition of CK2alpha' reduced ei activity by attenuation of hENT2 transcription, while the increase in es/hENT1 activity was mediated by post-translational action of CK2. The observed modification in es activity was probably due to a CK2alpha'-mediated change in the phosphorylation state of the ENT1 protein, or an interacting protein, effecting an increase in the plasma membrane lifetime of the transport proteins. |
| Databáze: | OpenAIRE |
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