Polymorphic kinase anchor proteins and nucleic acids encoding the same

Abstract

Polymorphic A-kinase anchor proteins (AKAPs) and nucleic acids encoding the proteins are provided herein. Methods of detecting polymorphic AKAPs and nucleic acids encoding the AKAPs, and kits for use in the detection methods are also provided. Further provided herein are methods of identifying subjects having or at risk of developing disorders of signal transduction. Methods of determining susceptibility to morbidity and/or increased or early mortality are also provided.

1 . A method for detecting the presence or absence of an allelic variant of a human AKAP10 gene, comprising determining the identity of the nucleotide at a position corresponding to position 2073 of the coding sequence of a human AKAP10 gene or the complement thereof, wherein a variant has a nucleotide other than A at a position corresponding to position 2073.  
     
     
         2 . The method of  claim 1 , wherein determining the identity comprises: 
 (a) hybridizing a target nucleic acid comprising a human AKAPIO-encoding nucleic acid or fragment thereof or a complement of a human AKAPIO-encoding nucleic acid or fragment thereof with a nucleic acid primer that hybridizes adjacent to a position corresponding to position 2073 of the coding sequence of the human AKAP10 gene or complement thereof;    (b) extending the nucleic acid primer using the target nucleic acid as a template; and    (c) determining the mass of the extended primer to identify the nucleotide present at a position corresponding to position 2073 or the complement thereof, thereby determining the presence or absence of an allelic variant.    
     
     
         3 . The method of  claim 2 , wherein the mass is determined by mass spectrometry.  
     
     
         4 . The method of  claim 2 , wherein the primer is extended in the presence of at least one dideoxynucleotide,  
     
     
         5 . The method of  claim 4 , wherein the at least one dideoxynucleotide is ddT.  
     
     
         6 . The method of  claim 4 , wherein the primer is extended in the presence at least two dideoxynucleotides and the at least two dideoxynucleotides are ddT and ddC.  
     
     
         7 . The method of  claim 6 , wherein the mass is determined by mass spectrometry.  
     
     
         8 . The method of  claim 4 , wherein the at least one dideoxynucleotide is ddA.  
     
     
         9 . The method of  claim 4 , wherein the primer is extended in the presence at least two dideoxynucleotides and the at least two dideoxynucleotides are ddA and ddG.  
     
     
         10 . The method of  claim 9 , wherein the mass is determined by mass spectrometry.  
     
     
         11 . The method of  claim 2 , wherein hybridization is effected under conditions of high stringency.  
     
     
         12 . The method of  claim 1 , wherein determining the identity comprises: 
 (a) hybridizing a target nucleic acid comprising a human AKAPIO-encoding nucleic acid, complement thereof or fragment thereof with a single-stranded nucleic acid probe at a position corresponding to position 2073 of the coding sequence of the human AKAP10 gene or complement thereof; and    (b) detecting hybridized probe to identify the nucleotide present at a position corresponding to position 2073 or the complement thereof, thereby determining the presence or absence of an allelic variant.    
     
     
         13 . The method of  claim 12 , wherein hybridization is effected under conditions of high stringency.  
     
     
         14 . The method of  claim 12 , wherein the nucleotide of the probe that hybridizes with the nucleotide at a position corresponding to position 2073 is complementary to a G, T, or C nucleotide.  
     
     
         15 . The method of  claim 12 , wherein the nucleotide of the probe that hybridizes with the nucleotide at the complement of a position corresponding to position 2073 is complementary to a G, A, or C nucleotide.  
     
     
         16 . The method of  claim 14 , wherein the nucleotide detected at the position corresponding to position 2073 is a G.  
     
     
         17 . The method of  claim 15 , wherein the nucleotide detected at the complement of the position corresponding to position 2073 is a C.  
     
     
         18 . A method for indicating susceptibility to morbidity, increased or early mortality, or morbidity and increased or early mortality of a subject, comprising: 
 detecting the presence or absence of at least one allelic variant of a polymorphic region of an AKAP10 gene that is associated with    susceptibility to morbidity, increased or early mortality, or morbidity and increased or early mortality, wherein:    the predominant allele comprises an “A” at a position corresponding to position 2073 of SEQ ID NO: 1; and    the presence of the allelic variant is indicative of increased susceptibility to morbidity, increased or early mortality, or morbidity and increased or early mortality compared to the susceptibility of a subject who does not comprise the allelic variant.    
     
     
         19 . The method of  claim 18 , wherein a polymorphic region of the AKAP10 gene comprises a nucleotide other than an A at a position corresponding to position 2073 of the coding sequence of the AKAP10 gene or other than an T of the complement of the coding sequence of the AKAP10 gene.  
     
     
         20 . The method of  claim 18 , wherein the detecting step is effected by a method selected from the group consisting of allele specific hybridization, primer specific extension, oligonucleotide ligation assay, restriction enzyme site analysis and single-stranded conformation polymorphism analysis.  
     
     
         21 . The method of  claim 18 , wherein the detecting step comprises mass spectrometry.  
     
     
         22 . The method of  claim 18 , wherein detection is effected by detecting a signal moiety selected from the group consisting of radioisotopes, enzymes, antigens, antibodies, spectrophotometric reagents, chemiluminescent reagents, fluorescent reagents and other light producing reagents.  
     
     
         23 . The method of  claim 19 , further comprising: 
 detecting an allelic variant at another polymorphic region of an AKAP10 gene selected from the group consisting of a position corresponding to position 83587 of SEQ ID NO: 13, a position corresponding to position 129,600 of SEQ-ID NO: 14 and a position corresponding to position 156,277 of SEQ ID NO: 18.    
     
     
         24 . A method for indicating an alteration in signal transduction in a subject, comprising: 
 detecting the presence or absence of an allelic variant of an AKAP10 gene having a nucleotide other than A at a position corresponding to position 2073 of SEQ ID NO: 1, wherein the presence of a nucleotide other than A is indicative of an alteration in signal transduction.    
     
     
         25 . The method of  claim 24 , wherein the allelic variant has a G at a position corresponding to position 2073 of SEQ ID NO: 1.  
     
     
         26 . The method of  claim 25 , further comprising detecting the presence or absence of an allelic variant at another polymorphic position of the AKAP10 gene selected from the group consisting of a position corresponding to position 83587 of SEQ ID NO: 13, position 129,600 of SEQ ID NO: 14 and position 156,277 of SEQ ID NO: 18.  
     
     
         27 . The method of  claim 24 , wherein the alteration in signal transduction is related to a disorder selected from the group consisting of cardiovascular disorders, cardiac disorders, proliferative disorders, 
 neurological disorders, neurodegenerative disorders, obesity, diabetes and peripheral retinopathies.    
     
     
         28 . The method of  claim 27 , wherein the disorders are selected from the group consisting of Alzheimer's disease, altered left ventricular function, cardiomyopathies, bipolar disorder and retinitis pigmentosa.  
     
     
         29 . The method of  claim 27 , wherein the detecting step is effected by a method selected from the group consisting of allele specific hybridization, primer specific extension, oligonucleotide ligation assay, restriction enzyme site analysis and single-stranded conformation polymorphism analysis.  
     
     
         30 . The method of  claim 24 , wherein the detecting step comprises mass spectrometry.  
     
     
         31 . The method of  claim 24 , wherein the detecting step is effected a signal moiety selected from the group consisting of: radioisotopes, enzymes, antigens, antibodies, spectrophotometric reagents, chemiluminescent reagents, fluorescent reagents and other light producing reagents.