Parkin Transcript Variants in Rat and Human Brain

Alternative splicing has an important role in expanding protein diversity. We have identified complementary DNA species from adult rat and fetal human brain encoding seven new splice variants of parkin, a gene mutated in autosomal recessive juvenile parkinsonism (ARJP). Alternative splicing affects almost all previously characterized exons, plus 3 new exons of 72, 156, and 180 nucleotides. This creates the potential to express hundreds of different isoforms. The encoded parkin isoforms have different amino acid composition, post-translational modifications, and, most important, molecular architectures. They diverge for the presence or absence of the ubiquitin-like domain, one or two C3HC4 ring fingers, the in-between ring fingers (IBR) domain, and a thiol proteases active site, which has not been previously characterized. Distinct expression patterns occur in primary cultures of neuronal and glial cells. Extensive splicing of parkin produces regional and structural diversity and may have important implications for the pathogenetic mechanisms underlying ARJP.     

Human Microtubule-Associated Protein 1a (MAP1A) Gene: Genomic Organization, cDNA Sequence, and Developmental- and Tissue-Specific Expression

Microtubule-associated proteins (MAPs) regulate microtubule stability and play critical roles in neuronal development and the balance between neuronal plasticity and rigidity. MAP1a (HGMW-approved symbol MAP1A) stabilizes microtubules in postnatal axons. We describe human MAP1a's genomic organization and deduced cDNA and amino acid sequences. MAP1a is a single-copy gene spanning 10.5 kb. MAP1a coding sequence is contained in five exons. Translation begins in exon 3. Human MAP1a contains 2805 amino acids (predicted molecular weight 306.5 kDa) and is slightly larger than rat MAP1a (2774 amino acids). Like rat and bovine MAP1a, human MAP1a contains conserved tubulin binding motifs in the amino-terminal region. The carboxy-terminal portion contains a conserved pentadecapeptide that is present in the light chain portion of rat and bovine MAP1a/LC2 polyprotein. We show that human MAP1a gene expression occurs almost exclusively in the brain and that there is approximately 10-fold greater gene expression in adult brain compared to fetal brain. Strong, interspecies conservation between human and rat MAP1a cDNA and amino acid sequences indicates important relationships between MAP1a's function and its primary amino acid sequence.     

Developmentally regulated alternative RNA splicing of rat brain sodium channel mRNAs.

Two rat brain Na channel alpha-subunit cDNAs, named RII and RIIA, have almost identical coding regions, with a divergence of only 36 nucleotides (0.6%) over a total length of 6015 residues. A cluster of 20 divergent residues occurs within a 90 nucleotide segment of cDNA sequence. We now demonstrate that this 90 nucleotide segment is encoded twice in the RII/RIIA genomic sequence. Furthermore, the mutually exclusive selection of these two exons is developmentally regulated. RII mRNAs are relatively abundant at birth but are gradually replaced by RIIA mRNAs as development proceeds. The two mRNAs also appear to have different regional distributions in the developing rat brain. Strikingly, although 30 amino acids are encoded by each alternative exon, only amino acid position 209 is altered between the two, specifying asparagine in RII and aspartate in RIIA. Alternative RNA splicing may modulate the RII/RIIA sodium channel properties during neuronal development.