Genomic organisation and chromosomal localisation of the gene encoding human beta adducin

doi:10.1016/0378-1119(95)00591-9

Gene

Volume 167, Issues 1–2, 29 December 1995, Pages 313-316

https://doi.org/10.1016/0378-1119(95)00591-9 Get rights and content

Abstract

Adducin (ADD) is a heterodimeric protein of the membrane skeleton with subunits of 103 (α) and 97 kDa (β). It promotes the assembly of the spectrin-actin network. We have previously shown that one point mutation in each of the α and β rat ADD-encoding genes is associated with blood pressure variation in an animal model for hypertension, the Milan hypertensive strain of rats, probably due to a change in the phosphorylation pattern. In fact, the rat mutations, Y to F for α and R to Q for β, are located, respectively, in a tyrosine kinase and a protein kinase A phosphorylation site. We have now determined, for the human β-ADD-encoding gene, its chromosomal localisation, exon-intron organisation and alternative splicing patterns. We report here that human β-ADD is localised on chromosome 2 and we also show a characteristic 3′ end alternative splicing of the β-ADD RNA that generates two distinct β-ADD families, namely ADD 63 and 97; both of them in turn present a very complex differential splicing pattern in the internal exons.

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Cited by (19)

Expression analysis of the human adducin gene family and evidence of ADD2 β 4 multiple splicing variants
2003, Biochemical and Biophysical Research Communications
Adducin is a cytoskeleton heterodimeric protein. Its subunits are encoded by three related genes (ADD1, ADD2, and ADD3) which show alternative spliced variants. Adducin polymorphisms are involved in blood pressure regulation in humans and rats. We have analyzed mRNA distribution of ADD gene family in human tissues and cells with Real-Time TaqMan RT-PCR. Whereas ADD1 is ubiquitously distributed, ADD3 is more expressed in kidney medulla and cortex than in fetal kidney, while in adult liver it is less abundant than in fetal liver. ADD2 β1 and β4 variants show the same pattern of distribution with the highest expression in brain, fetal liver, and kidney. Conventional RT-PCR identified new β4 variants. β4a is characterized by an in-frame insertion of 21 nucleotides upstream exon 15 predicting a 7 amino acids longer protein with a similar C-terminus region. It is coexpressed with β1 and β4 in several tissues. Fetal kidney shows further β4b, β4c and β4d variants containing internal exon deletions that enormously modify the predicted NH₂ and central regions. Our findings could help one to understand the functional role of adducin variants in specific tissues and cells.
Utilization of an 86 bp exon generates a novel adducin isoform (β4) lacking the MARCKS homology domain
1998, Biochimica et Biophysica Acta - Gene Structure and Expression
A novel isoform of β-adducin has been amplified and characterized from a human bone marrow cDNA library (GenBank #U43959). This isoform arises from the insertion of an 86 bp alternatively spliced and previously unrecognized exon (now termed exon 15) within codon 581 of the human red blood cell β-adducin sequence. This results in an insertion of 28 novel amino acids. The remainder of the red cell β-adducin mRNA is then translated in a different reading frame, adding an additional 35 novel amino acids prior to the stop codon. This new isoform, thus, replaces β1-adducin sequence after residue 580 with a total of 63 new amino acids. Sequences from genomic clones of the human β-adducin gene show that this alternate exon is flanked by splice consensus sequences and is appropriately located in the genomic map between exons encoding up-stream and down-stream sequences, thus defining a new exon. The COOH-terminus of this new isoform, which we designate β4, lacks a 22 amino acid lysine-rich sequence common to both the human red cell α- and β-adducin subunits and homologous to a highly conserved region in MARCKS, a filamentous actin-cross linking protein regulated by protein kinase C and calcium /calmodulin. β4-adducin preserves a previously identified calmodulin binding domain. PCR analysis indicates that this new β-adducin isoform is expressed in fetal brain and liver, bone marrow, and NT-2 (neuroepithelial) cells, but is not detected in several other tissues. We anticipate that this new β4 isoform of β-adducin will display unique and tissue-specific functional properties.
Organization of the human β-adducin gene (ADD2)
1997, Genomics
The intron–exon organization of the human β-adducin gene (ADD2) has been determined from overlapping genomic clones. The gene spans over 100 kb on chromosome 2p13 and comprises 17 exons. Seven of the exons are identical in size to the corresponding exons of the α-adducin gene (4p16.3), suggesting gene duplication. A 275-bp fragment 5′ to exon 1 demonstrates strong promoter activity in a transient transfection assay. Within 333 bp 5′ of the first exon can be found several putative transcription factor-binding sites: three SP1 sites, one GATA site, three MZF1 sites, one p300 site, and one c-Ets site. Alternatively spliced exons in the 3′ region are described and contain distinct coding regions, stop codons, and 3′UTR, corresponding to previously published β-adducin cDNA sequences β-1 and β-2. The alternative splice sites for the smallest adducin isoform, β-3, are alternative donor and acceptor sites within exons 7 and 12. The most recently described isoform, β-4, includes an alternative exon (exon 15) that results in a frame shift and early termination. Intron–exon splice sites are presented for all 17 exons and conform to the consensus sequences for mammalian splice sites. These results will be useful in further analysis of tissue-specific expression of adducin isoforms and in analysis of DNA from patients with diseases mapping to this region of chromosome 2.
Characterisation and chromosomal localisation of the rat α- and β-adducin-encoding genes
1995, Gene
A polymorphism in the genes encoding α- and β-adducin (ADD) was described as being associated with blood-pressure variation in a genetically hypertensive strain of rats (MHS). ADD is a cytoskeletal heterodimeric protein which may be involved in cellular signal transduction and interacts with other membrane skeleton proteins which affect ion transport across the cell membrane. The cDNA encoding the α- subunit of rat ADD was isolated using PCR methods. The cDNA consists of about 3900 bp and encodes a protein of 735 amino acids (aa) which shows 91% aa identity with the human counterpart. In spleen and kidney, three alternative spliced exons were found by PCR amplification and confirmed by RNase protection analysis. 17 inbred rat strains were genotyped for the polymorphism in the α- and β-ADD genes. Chromosomal localisation mapped rat α-ADD on chromosome 14 and rat β-ADD on chromosome 4.
The Association between Gly460Trp-Polymorphism of Alpha-Adducin 1 Gene (ADD1) and Arterial Hypertension Development in Ukrainian Population
2021, International Journal of Hypertension
A review of the epidemiological evidence for adducin family gene polymorphisms and hypertension
2019, Cardiology Research and Practice

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Short communicationGenomic organisation and chromosomal localisation of the gene encoding human beta adducin

Abstract

J. Biol. Chem.

J. Biol. Chem.

Biochem. Biophys. Res. Commun.

Two point mutations within the adducin genes are involved in blood pressure variation

Cloning and mapping of the alpha-adducin gene close to D4S95 and assessment of its relationship to Huntington disease

Hum. Mol. Genet.

Localisation of single copy DNA sequences of G-banded human chromosomes by in situ hybridization

Chromosoma

Mapping the domain structure of human erythrocyte adducin

J. Biol. Chem.

Short communication
Genomic organisation and chromosomal localisation of the gene encoding human beta adducin