ISOLATION AND CHARACTERIZATION OF CDNAS ENCODING HUMAN BRAIN ANKYRINS REVEAL A FAMILY OF ALTERNATIVELY SPLICED GENES

被引:134
作者
OTTO, E [1 ]
KUNIMOTO, M [1 ]
MCLAUGHLIN, T [1 ]
BENNETT, V [1 ]
机构
[1] DUKE UNIV,MED CTR,DEPT BIOCHEM,DURHAM,NC 27710
关键词
D O I
10.1083/jcb.114.2.241
中图分类号
Q2 [细胞生物学];
学科分类号
071009 ; 090102 ;
摘要
Ankyrins are a family of membrane-associated proteins that can be divided into two immunologically distinct groups: (a) erythrocyte-related isoforms (ankyrin(R)) that have polarized distributions in particular cell types; and (b) brain-related isoforms (ankyrin(B)) that display a broader distribution. In this paper, we report the isolation and sequences of cDNAs related to two ankyrin(B) isoforms, human brain ankyrin 1 and 2, and show that these isoforms are produced from alternatively spliced mRNAs of a single gene. Human brain ankyrin 1 and 2 share a common NH2-terminus that is similar to human erythrocyte ankyrins, with the most striking conservation occurring between areas composed of a repeated 33-amino acid motif and between areas corresponding to the central portion of the spectrin-binding domain. In contrast, COOH-terminal sequences of brain ankyrin 1 and 2 are distinct from one another and from human erythrocyte ankyrins, and thus are candidates to mediate protein interactions that distinguish these isoforms. The brain ankyrin 2 cDNA sequence includes a stop codon and encodes a polypeptide with a predicted molecular mass of 202 kD, which is similar to the M(r) of the major form of ankyrin in adult bovine brain membranes. Moreover, an antibody raised against the conserved NH2-terminal domain of brain ankyrin crossreacts with a single M(r) = 220 kD polypeptide in adult human brain. These results strongly suggest that the amino acid sequence of brain ankyrin 2 determined in this report represents the complete coding sequence of the major form of ankyrin in adult human brain. In contrast, the brain ankyrin 1 cDNAs encode only part of a larger isoform. An immunoreactive polypeptide of M(r) = 440 kD, which is evident in brain tissue of young rats, is a candidate to be encoded by brain ankyrin 1 mRNA. The COOH-terminal portion of brain ankyrin 1 includes 15 contiguous copies of a novel 12-amino acid repeat. Analysis of DNA from a panel of human/rodent cell hybrids linked this human brain ankyrin gene to chromosome 4. This result, coupled with previous reports assigning the human erythrocyte ankyrin gene to chromosome 8, demonstrates that human brain and erythrocyte ankyrins are encoded by distinct members of a multigene family.
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页码:241 / 253
页数:13
相关论文
共 39 条
[1]   CLONING, SEQUENCING AND TRANSCRIPTIONAL CONTROL OF THE SCHIZOSACCHAROMYCES-POMBE-CDC10 START GENE [J].
AVES, SJ ;
DURKACZ, BW ;
CARR, A ;
NURSE, P .
EMBO JOURNAL, 1985, 4 (02) :457-463
[2]  
Bennett V, 1982, Cold Spring Harb Symp Quant Biol, V46 Pt 2, P647
[4]  
BENNETT V, 1983, METHOD ENZYMOL, V96, P313
[5]   SPECTRIN-BASED MEMBRANE SKELETON - A MULTIPOTENTIAL ADAPTER BETWEEN PLASMA-MEMBRANE AND CYTOPLASM [J].
BENNETT, V .
PHYSIOLOGICAL REVIEWS, 1990, 70 (04) :1029-1065
[6]   SIMILARITY BETWEEN CELL-CYCLE GENES OF BUDDING YEAST AND FISSION YEAST AND THE NOTCH GENE OF DROSOPHILA [J].
BREEDEN, L ;
NASMYTH, K .
NATURE, 1987, 329 (6140) :651-654
[7]   ROTATIONAL AND LATERAL DIFFUSION OF MEMBRANE-PROTEINS [J].
CHERRY, RJ .
BIOCHIMICA ET BIOPHYSICA ACTA, 1979, 559 (04) :289-327
[8]   XOTCH, THE XENOPUS HOMOLOG OF DROSOPHILA-NOTCH [J].
COFFMAN, C ;
HARRIS, W ;
KINTNER, C .
SCIENCE, 1990, 249 (4975) :1438-1441
[9]  
DAVIS J, 1989, J BIOL CHEM, V264, P6417
[10]  
DAVIS JQ, 1984, J BIOL CHEM, V259, P3550