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Sequence and Functional Conservation of the Intergenic Region Between the Head-to-Head Genes Encoding the Small Heat Shock Proteins αB-Crystallin and HspB2 in the Mammalian Lineage

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Abstract

An unexpected feature of the large mammalian genome is the frequent occurrence of closely linked head-to-head gene pairs. Close apposition of such gene pairs has been suggested to be due to sharing of regulatory elements. We show here that the head-to-head gene pair encoding two small heat shock proteins, αB-crystallin and HspB2, is closely linked in all major mammalian clades, suggesting that this close linkage is of selective advantage. Yet αB-crystallin is abundantly expressed in lens and muscle and in response to a heat shock, while HspB2 is abundant only in muscle and not upregulated by a heat shock. The intergenic distance between the genes for these two proteins in mammals ranges from 645 bp (platypus) to 1069 bp (opossum), with an average of about 900 bp; in chicken the distance was the same as in duck (1.6 kb). Phylogenetic footprinting and sequence alignment identified a number of conserved sequence elements close to the HspB2 promoter and two farther upstream. All known regulatory elements of the mouse αB-crystallin promoter are conserved, except in platypus and birds. The lens-specific region 1 (LSR1) and the heat shock elements (HSEs) lack in birds; in platypus the LSR1 is reduced to a Pax-6 site, while the Pax-6 site in LSR2 and a HSE are absent. Most likely the primordial mammalian αB-crystallin promoter had two LSRs and two HSEs. In transfection experiments the platypus αB-crystallin promoter retained heat shock responsiveness and lens expression. It also directed lens expression in Xenopus laevis transgenes, as did the HspB2 promoter of rat or blind mole rat. Deletion of the middle of the intergenic region including the upstream enhancer affected the activity of both the rat αB-crystallin and the HspB2 promoters, suggesting sharing of the enhancer region by the two promoters.

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Acknowledgments

We thank Erik Jansen for technical support with the Xenopus transgenesis. This investigation was supported by the Research Council for Earth and Life Sciences (ALW) with financial aid from the Netherlands Organization for Scientific Research (NWO) and by the European Commission (TMR ERB-FMRX-CT98-0221 and BMH4-CT98-3895).

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Authors and Affiliations

  1. Department of Biochemistry, Faculty of Science, University of Nijmegen, Nijmegen, The Netherlands

    Linda Doerwald, Teun van Rheede, Ole Madsen, Remco Rexwinkel, Siebe T. van Genesen, Wilfried W. de Jong & Nicolette H. Lubsen

  2. Department of Animal Physiology, Faculty of Science, University of Nijmegen, Nijmegen, The Netherlands

    Ron P. Dirks & Gerard J. Martens

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  1. Linda Doerwald
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  2. Teun van Rheede
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  5. Remco Rexwinkel
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  6. Siebe T. van Genesen
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  9. Nicolette H. Lubsen
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Correspondence to Nicolette H. Lubsen.

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Reviewing Editor: Dr. Manyuan Long

(Linda Doerwald and Teun Van Rheede) Both authors contributed equally.

(Teun van Rheede) Deceased May 21, 2003.

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Doerwald, L., van Rheede, T., Dirks, R.P. et al. Sequence and Functional Conservation of the Intergenic Region Between the Head-to-Head Genes Encoding the Small Heat Shock Proteins αB-Crystallin and HspB2 in the Mammalian Lineage. J Mol Evol 59, 674–686 (2004). https://doi.org/10.1007/s00239-004-2659-y

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