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Volume 272, Number 15, Issue of April 11, 1997 pp. 10013-10020
©1997 by The American Society for Biochemistry and Molecular Biology, Inc.

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Cloning and Chromosomal Location of a Novel Member of the Myotonic Dystrophy Family of Protein Kinases

(Received for publication, August 30, 1996, and in revised form, December 12, 1996)

Yanming Zhao , Pascal Loyer ^¶ , Haimin Li ^¶ , Virginia Valentine ^** , Vincent Kidd ^¶ and Andrew S. Kraft

From the  Division of Medical Oncology, University of University of Colorado Health Sciences Center, Denver, Colorado 802621 and the Departments of ^¶ Tumor Cell Biology and ^** Experimental Oncology, St. Jude Children's Hospital, Memphis, Tennessee 38105

We have cloned a novel serine/threonine protein kinase (PK428) which is highly related (65%) within the kinase domain to the myotonic dystrophy protein kinase (DM-PK), as well as the cyclic AMP-dependent protein kinase (33%). Northern blots demonstrate that PK428 mRNA is distributed widely among tissues and is expressed at the highest levels in pancreas, heart, and skeletal muscle, with lower levels in liver and lung. Two PK428 mRNAs 10 and 3.8 kilobase pairs in size are seen in a number of cell lines, including hematopoietic and breast cancer cells. An antibody generated to a glutathione S-transferase-PK428 fusion protein detects a 65-kDa protein in these cell lines, and a similarly sized protein when the cloned cDNA is transiently expressed in Cos 7 cells. Immunoprecipitation of the transiently expressed PK428 protein and incubation with [-³²P]ATP demonstrate that it is capable of autophosphorylation. In addition, immunoprecipitates of the PK428 protein kinase also phosphorylated histone H1 and a peptide encoding a cyclic AMP-dependent protein kinase substrate. The gene corresponding to the 3.8-kb PK428 mRNA, and its corresponding 65-kDa protein, was isolated by polymerase chain reaction screening of a P1 phage human genomic library. Using this P1 phage clone as a probe, the PK428 gene was located on 1q41-42, a possible location for a human senescence gene, a gene associated with Rippling muscle disease, as well as a region associated with genetically acquired mental retardation.

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