Abstract

We report the determination of the complete DNA sequence for c beta 3, a chicken beta-tubulin gene which we show to be the dominant beta-tubulin expressed in testis. Like all previously studied vertebrate beta-tubulin genes, the gene is divided into four exon sequences interrupted by three intervening sequences (located between amino acids 19 and 20, within codon 56, and within codon 93). Analysis of the program of expression of this gene indicates that it encodes the dominant chicken testis beta-tubulin, although it is also expressed at lower levels in a wide variety of cell and tissue types. Comparison of the predicted polypeptide sequence for c beta 3 with four other available chicken beta-tubulin genes confirms our earlier suggestion that within an otherwise conserved framework, sequences within two variable region domains serve to define specific beta-tubulin polypeptide isotypes. The data indicate that the c beta 3 gene encodes a unique beta-tubulin isotype which diverges from the dominant neuronal beta-tubulin isotype in 18 of 445 residues (4%). Although the protein coding regions of the c beta 3 gene are highly homologous to the chicken c beta 1, c beta 2, c beta 4, and c beta 5 genes previously reported by us, no significant sequence homology with these previously analyzed genes is discernible in the 5'- or 3'-untranslated region sequences, in the intervening sequences, or in the presumptive transcriptional promoter sequences.

Highlights

  • We report the determination of the complete DNA processes [4,5].Microtubules play a centralrole in other sequence for cS3, a chicken &tubulin gene which we forms of intracellular transport [6] and in transduction of show tobe the dominant @-tubulin expressed tinestis. signals originating at thecell membrane; and, asmajor struc

  • Analysis of the program ofexpression ofthis gene indicates thatit encodes the dominant chicken testis #?-tubulin, althoughit is expressed at lower levels inawide variety of cell and tissue types.Comparisonofthepredictedpolypeptide seare encoded in vertebrate organisms by small multigene families

  • The dataalso indicate that thec@3gene encodes a unique chicken @-tubulinisotype which diverges from the dominant neuronal @-tubulinisotype

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Summary

A VERTEBRATE TESTIS B-TUBULIN ISOTUPE*

From the D e ~ r ~ ~ofn~ t ~s ~ oCh~emigstry~anad fllCeU 3 ~The J~ohns ~~ o,~ n ~ u~e T sSic~h~yool of ~i e d ~~ i ~ , att ti more, ~ arytan d21205. Different @-tubulingenes contribute to differential microtubule functions might unfold from such an analysis To this end, we have determined the complete DNA sequence for c@3, a chicken @-tubulingene which we show to be the dominant @-tubulinexpressed in testis. Radiolabeled DNA probes were prepared using (a-32P]dATPand therandom priming method of Shank et al [29], as previously described [8].All untranslated regions of the gene, the transcription initiation site for 12/33was determined by using S1 nuclease analysis as describedunder "Materials anMd ethods." ,a fragment from c83 that begins at nucleotide 12 with respect to the A of the ATG translation initiation codon was labeled at probes were labeled to approximately equal specific activities, with the 5' ends andwas used to probe poly(A)+RNA from chicken the exception of 82-3' which gave %fold lower activity. Probe fragments protected from digestion as the result of hybridization to c83 RNAs were electrophoresed ona6%

RESULTS
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DISCUSSION
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