Targeting of a Distinctive Protein-Serine Phosphatase to the Protein Kinase-Like Domain of the Atrial Natriuretic Peptide Receptor

• Published in: Proceedings of the National Academy of Sciences - PNAS Vol. 91; no. 23; pp. 11075 - 11079
• Main Author: Chinkers, Michael
• Format: Journal Article
• Language: English
• Published: United States National Academy of Sciences of the United States of America 08.11.1994; National Acad Sciences; National Academy of Sciences
• Subjects: Amino Acid Sequence; Animals; Cells; Cellular biology; Cloning, Molecular; Complementary DNA; Embryos; Gene Expression; Guanylate Cyclase - metabolism; Libraries; Mathematical functions; Mice; Molecular Sequence Data; Phosphatases; Phosphoprotein Phosphatases - metabolism; Plasmids; Protein Binding; Protein Kinases - chemistry; Proteins; Rats; Receptors; Receptors, Atrial Natriuretic Factor - metabolism; Recombinant Proteins - metabolism; RNA, Messenger - genetics; Sequence Alignment; Sequence Homology, Amino Acid; Signal Transduction; Yeasts
• ISSN: 0027-8424; 1091-6490
• DOI: 10.1073/pnas.91.23.11075

Protein kinase-related domains of unknown function are present in the JAK family of protein tyrosine kinases and in receptor/guanylyl cyclases. I used the yeast two-hybrid system to screen for proteins interacting with the kinase-like domain of the atrial natriuretic peptide (ANP) receptor/guanylyl cyclase. A yeast strain was constructed expressing a fusion of this kinase-like domain to the lexA DNA-binding domain and containing a HIS3 gene under the control of lexA upstream activating sequences. These yeast cells were transformed with a plasmid library of mouse embryo cDNA fragments fused to the VP16 transcriptional activation domain. Cells containing VP16-fusion proteins interacting with the lexA-kinase-like domain fusion protein were selected by growth in the absence of histidine. A partial-length cDNA clone isolated by using this approach encoded a protein that interacted specifically with the ANP-receptor protein kinase-like domain both in yeast cells and in vitro. Tissue-specific expression of a 2.2-kb mRNA hybridizing to this cDNA paralleled the known pattern of ANP-receptor mRNA expression. A full-length cDNA clone isolated from a rat lung library was predicted to encode a 55-kDa protein containing at its amino terminus a targeting domain that binds to the ANP-receptor kinase-like domain and containing at its carboxyl terminus a putative protein-serine phosphatase domain. This protein is a possible candidate for the phosphatase involved in desensitizing the ANP receptor. Targeting of regulatory proteins may be an important function of protein kinase-like domains.