- Email: [email protected]
Alternative splicing and structure of the human erythroid dematin gene
Biochimica et Biophysica Acta 1398 (1998) 382^386
Short sequence-paper
Alternative splicing and structure of the human erythroid dematin gene Anthony C. Kim, Anser C. Azim 1 , Athar H. Chishti * Laboratory of Tumor Cell Biology, St. Elizabeth's Medical Center, Tufts University School of Medicine, ACH4, 736 Cambridge Street, Boston, MA 02135, USA Received 23 February 1998; revised 15 April 1998; accepted 22 April 1998
Abstract Human erythroid dematin is a cytoskeletal protein capable of bundling actin filaments in vitro. The carboxyl terminal domain of dematin is homologous to the headpiece domain of villin, an actin-binding protein of the brush border cytoskeleton. Here we report the complete structure of the dematin gene located on human chromosome 8p21.1, a region frequently deleted in prostate cancer. The dematin gene is composed of 15 exons spanning V15 kb. We also report two novel isoforms of dematin derived from alternative splicing of the dematin gene in the brain. ß 1998 Elsevier Science B.V. All rights reserved. Keywords: Dematin; Erythroid cytoskeleton; Headpiece; Actin-binding; Tumor suppressor gene; Prostate cancer
Dematin is an actin bundling phosphoprotein of the erythroid membrane cytoskeleton [1]. The primary structure of dematin is organized into an Nterminal domain and a C-terminal domain that is homologous to the `headpiece' domain of villin, an actin-binding protein of the brush border cytoskeleton [1]. In solution, erythroid dematin exists as a trimer of two 48 kDa subunits and one 52 kDa subunit [2]. We have recently cloned a protein with high homology to dematin's N-terminal and headpiece domains. Termed limatin (abLIM), this protein also contains four LIM domains at its amino-terminus and is the closest homolog of dematin known to date [3,4]. The core of the erythroid cytoskeleton consists of * Corresponding author. Fax: +1 (617) 789-3111; E-mail: [email protected] 1 Present address: Division of Experimental Medicine, Brigham and Women's Hospital, Harvard Medical School, Boston, MA 02115, USA.
spectrin, ankyrin, protein 4.1, and dematin [5]. While much is known about the functions of these proteins, little is known concerning the physiological function of dematin in vivo. In vitro, dematin bundles actin ¢laments in a phosphorylation-dependent manner [6]. While dematin is a substrate of multiple protein kinases, only phosphorylation by the cAMP-dependent protein kinases abolishes its actin-bundling activity [6]. Currently, we are investigating the function of dematin in non-erythroid cells. Dematin transcripts are widely expressed, particularly in the heart, brain, and skeletal muscle [1]. We have also mapped the locus encoding the dematin gene to human chromosome 8p21.1 [2]. In a review of the literature, the chromosomal region 8p21.1 is often deleted in a variety of cancers, including prostate cancer, which suggests the presence of a tumor suppressor gene(s) [7,8]. Here we report the complete structure of the human dematin gene. The availability of dematin's intron^exon organization will be useful for the analysis of possible genetic abnormalities of the dematin
0167-4781 / 98 / $19.00 ß 1998 Elsevier Science B.V. All rights reserved. PII: S 0 1 6 7 - 4 7 8 1 ( 9 8 ) 0 0 0 7 8 - 5
BBAEXP 91146 1-7-98
A.C. Kim et al. / Biochimica et Biophysica Acta 1398 (1998) 382^386
383
Fig. 1. Nucleotide sequence of the intron^exon boundaries of the dematin gene. Nucleotide positions of the intron^exon boundaries correspond to the 52 kDa dematin cDNA (accession no. U28389).
gene in human cancers. We also report the ¢nding of two novel isoforms of dematin in the brain, both lacking exon 2. We used a previously isolated P1 clone to determine the genomic structure of the dematin gene [2]. P1#77 was digested with KpnI and shotgun sub-
cloned into the vector pGEM-4Z. Two of the KpnI subclones contained the entire dematin gene by PCR analysis. Intron^exon boundaries were determined by sequencing the KpnI subclones using primers derived from the dematin cDNA (Fig. 1). Intron sizes were determined by a combination of PCR and DNA
Fig. 2. Genomic organization of the human dematin gene. (A) Full-length erythroid dematin cDNA (52 kDa). (B) Dematin isoforms in erythrocytes and brain. (C) The intron^exon map of the human dematin gene derived from P1 genomic clone #77. Exons 2 and 13 (¢lled black) are alternative coding exons.
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Fig. 3. Expression of dematin mRNA in various tissues. Dematin is primarily expressed as two transcripts of 1.35 and 2.5 kb in length. The 2.5 kb transcript is particularly abundant in the prostate (lane 3), testis (lane 4), and small intestines (lane 6). The lower panel shows the same blot hybridized with L-actin to normalize for loading di¡erences.
sequence analysis. The dematin gene consists of 15 exons interrupted by 14 introns and spans approximately 15 kb (Fig. 2). Exon 13 encodes an alternatively spliced 22 amino acid insertion found in the 52 kDa subunit of erythroid dematin [2]. The PEST sequence, a motif found in signal transduction proteins that undergo rapid turnover in vivo, spans the boundary between exons 4 and 5. A negatively charged cluster of 10 glutamic and aspartic acid residues, a motif commonly found in proteins associated with the cell nucleus, is encoded by exon 8.
We used the BLAST 2.0 program to query the dematin cDNA against the human expressed sequence tags (ESTs) database at the National Center for Biotechnology Information (NCBI) website [9]. Two ESTs (accession nos. H14221 and R87550) matched perfectly with dematin and contained identical gaps in their sequence. Both EST sequences are derived from the 5P end of dematin cDNAs cloned from a human brain cDNA library. Complete DNA sequencing of the cDNAs revealed both to be novel full-length isoforms of dematin. EST R87550 encodes a cDNA that lacks exon 2 while EST H14221 lacks exons 2 and 13 (Fig. 2). The functional signi¢cance of dematin transcripts lacking exon 2 or exons 2 and 13 is presently unknown. Sequence analysis of the 25 amino acids that exon 2 encodes reveals no obvious homologies to any sequences deposited in GenBank. To analyze the expression of dematin in non-erythroid cells, we used the full-length dematin cDNA to probe the Human Multiple Tissue Northern Blot II using standard techniques (Clontech). Each lane is loaded with V2 Wg poly A RNA (Fig. 3). While villin is expressed exclusively in the brush border of intestinal and kidney cells, dematin transcripts are widely expressed, particularly so in the prostate and intestinal tissues. Two major transcripts of V1.35 and 2.5 kb in length hybridized to the cDNA probe. The 2.5 kb message is the predominant transcript in all tissues except in the thymus, and is consistent in size with the reported dematin full-length cDNA [1]. The cloning of dematin revealed it to be the ¢rst protein with sequence homology to the C-terminal headpiece domain of villin [1]. Since then, several proteins have been identi¢ed with similar modules,
Fig. 4. Amino acid sequence alignment of the headpiece domains of human dematin, villin, and limatin. Alignment was performed using the MegAlign software (DNAStar). Villin lacks the 22 amino acid insertion found in dematin and limatin.
BBAEXP 91146 1-7-98
A.C. Kim et al. / Biochimica et Biophysica Acta 1398 (1998) 382^386
including limatin and supervillin [3,10]. The dematin headpiece displays 32% amino acid identity with villin and 53% identity with limatin (Fig. 4). Interestingly, limatin is the only headpiece-containing protein to also contain the 22 amino acid insertion found in the 52 kDa dematin headpiece. The dematin headpiece insertion contains the phosphate binding consensus sequence GxGxxGR (P-loop) and has been shown to bind ATP in vitro [11]. Whether limatin similarly binds to nucleotides via the headpiece insertion in currently unknown. However, nucleotide binding seems unlikely since the limatin insertion varies considerably from the P-loop consensus sequence. Previously we, along with other groups, have demonstrated that the headpiece domain binds to actin ¢laments [2,3,12]. However, there is some controversy concerning the physiological function of the headpiece domain in vivo. While transfection studies of mutant villin cDNAs revealed the headpiece to be crucial in the morphogenesis of microvilli, recent evidence from knockout mice suggests that villin plays a minor or redundant role in the development of microvilli in absorptive tissues [12,13]. These studies raise the possibility that in villin null mice, dematin and/or other headpiece-containing proteins may substitute for villin in the reorganization of actin ¢laments in the absorptive epithelia. The short arm of chromosome 8 has been a region of intense scrutiny because of its propensity to undergo deletion in a variety of human cancers [7,8]. Recent evidence has been mounting implicating cytoskeletal proteins as tumor suppressors. Altered expression of the actin-binding proteins tropomyosin, gelsolin, K-actinin, and vinculin suppress the tumorigenesis of epithelial cells [14]. The brain tumor suppressor merlin/schwannomin, a member of the cytoskeletal protein 4.1 superfamily, suppresses the growth of NIH 3T3 cells [15]. The location of the dematin gene at 8p21.1 suggests that dematin may similarly be involved in the regulation of cell proliferation via its ability to organize actin bundles. In fact, we have recently observed the loss of one allele of the dematin gene in a majority of 8p21-linked prostate tumors [16]. Now that the intron^exon structure of the dematin gene has been elucidated, it will be possible to analyze whether the dematin gene is genetically altered in human cancers display-
385
ing loss of heterozygosity of chromosome 8p21.1. It is intriguing to note here that the limatin gene is located at chromosome 10q25, another region frequently deleted in human cancers, including prostate cancer [4]. This work was supported by NIH Grants HL51445 and CA66263 (A.H.C.). A.H.C. is an Established Investigator of the American Heart Association.
References [1] A.P. Rana, P. Ru¡, G.J. Maalouf, D.W. Speicher, A.H. Chishti, Cloning of human erythroid dematin reveals another member of the villin family, Proc. Natl. Acad. Sci. USA 90 (1993) 6651^6655. [2] A.C. Azim, J.H.M. Knoll, A.H. Beggs, A.H. Chishti, Isoform cloning, actin binding, and chromosomal localization of human erythroid dematin, a member of the villin superfamily, J. Biol. Chem. 270 (1995) 17407^17413. [3] D.J. Roof, A. Hayes, M. Adamian, A.H. Chishti, T. Li, Molecular characterization of abLIM, a novel actin-binding and double zinc ¢nger protein, J. Cell Biol. 138 (1997) 575^ 588. [4] A.C. Kim, L. L Peters, J.H.M. Knoll, C. Van Hu¡el, S.L. Ciciotte, P.W. Kleyn, A.H. Chishti, Limatin (LIMAB1), an actin-binding LIM protein, maps to mouse Chromosome 19 and human Chromosome 10q25, a region frequently deleted in human cancers, Genomics 46 (1997) 291^293. [5] D.L. Siegel, D. Branton, Partial puri¢cation and characterization of an actin-binding protein, band 4.9, from human erythrocytes, J. Cell Biol. 100 (1985) 775^785. [6] A.H. Chishti, A. Levin, D. Branton, Abolition of actin-bundling by phosphorylation of human erythrocyte protein 4.9, Nature 334 (1988) 718^721. [7] S.F. Huang, S. Xiao, A.A. Renshaw, K.R. Loughlin, T.J. Hudson, J.A. Fletcher, Fluorescence in situ hybridization evaluation of chromosome deletion patterns in prostate cancer, Am. J. Pathol. 149 (1996) 1565^1573. [8] C.D. Vocke, R.O. Pozzatti, D.G. Bostwick, C.D. Florence, S.B. Jennings, S.E. Strup, P.H. Duray, L.A. Liotta, M.R. Emmert-Buck, W.M. Linehan, Analysis of 99 microdissected prostate carcinomas reveals a high frequency of allelic loss on chromosome 8p12-21, Cancer Res. 56 (1996) 2411^ 2416. [9] S.F. Altschul, T.L. Madden, A.A. Scha¡er, J. Zhang, Z. Zhang, W. Miller, D.J. Lipman, Gapped BLAST and PSIBLAST: a new generation of protein database search programs, Nucleic Acids Res. 25 (1997) 3389^3402. [10] K.N. Pestonjamasp, R.K. Pope, J.D. Wulfkuhle, E.J. Luna, Supervillin (p205): a novel membrane-associated, F-actinbinding protein in the villin/gelsolin superfamily, J. Cell Biol. 139 (1997) 1255^1269.
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[11] A.C. Azim, S.M. Marfatia, C. Korsgren, E. Dotimas, C.M. Cohen, A.H. Chishti, Human erythrocyte dematin and protein 4.2 (Pallidin) are ATP binding proteins, Biochemistry 35 (1996) 3001^3006. [12] E. Friederich, K. Vancompernolle, C. Huet, M. Goethals, J. Finidori, J. Vandekerckhove, D. Louvard, An actin-binding site containing a conserved motif of charged amino acid residues is essential for the morphogenesis e¡ect of villin, Cell 70 (1992) 81^92. [13] K.I. Pinson, L. Dunbar, L. Samuelson, D.L. Gumucio, Targeted disruption of the mouse villin gene does not impair
the morphogenesis of microvilli, Dev. Dyn. 211 (1998) 109^121. [14] A. Ben-Ze'ev, The cytoskeleton of cancer cells, Biochim. Biophys. Acta 780 (1995) 197^212. [15] M. Lutchman, G.A. Rouleau, The neuro¢bromatosis type 2 gene product schwannomin, suppresses growth of NIH 3T3 cells, Cancer Res. 55 (1995) 2270^2274. [16] A.C. Kim, M. Lutchman, S. Pack, Z.P. Zhuang, A.H. Chishti, Overexpression of dematin alters cell morphology of normal and prostate tumor epithelial cells, Mol. Biol. Cell Suppl. 8 (1997) 276a.
BBAEXP 91146 1-7-98
Short sequence-paper
Alternative splicing and structure of the human erythroid dematin gene Anthony C. Kim, Anser C. Azim 1 , Athar H. Chishti * Laboratory of Tumor Cell Biology, St. Elizabeth's Medical Center, Tufts University School of Medicine, ACH4, 736 Cambridge Street, Boston, MA 02135, USA Received 23 February 1998; revised 15 April 1998; accepted 22 April 1998
Abstract Human erythroid dematin is a cytoskeletal protein capable of bundling actin filaments in vitro. The carboxyl terminal domain of dematin is homologous to the headpiece domain of villin, an actin-binding protein of the brush border cytoskeleton. Here we report the complete structure of the dematin gene located on human chromosome 8p21.1, a region frequently deleted in prostate cancer. The dematin gene is composed of 15 exons spanning V15 kb. We also report two novel isoforms of dematin derived from alternative splicing of the dematin gene in the brain. ß 1998 Elsevier Science B.V. All rights reserved. Keywords: Dematin; Erythroid cytoskeleton; Headpiece; Actin-binding; Tumor suppressor gene; Prostate cancer
Dematin is an actin bundling phosphoprotein of the erythroid membrane cytoskeleton [1]. The primary structure of dematin is organized into an Nterminal domain and a C-terminal domain that is homologous to the `headpiece' domain of villin, an actin-binding protein of the brush border cytoskeleton [1]. In solution, erythroid dematin exists as a trimer of two 48 kDa subunits and one 52 kDa subunit [2]. We have recently cloned a protein with high homology to dematin's N-terminal and headpiece domains. Termed limatin (abLIM), this protein also contains four LIM domains at its amino-terminus and is the closest homolog of dematin known to date [3,4]. The core of the erythroid cytoskeleton consists of * Corresponding author. Fax: +1 (617) 789-3111; E-mail: [email protected] 1 Present address: Division of Experimental Medicine, Brigham and Women's Hospital, Harvard Medical School, Boston, MA 02115, USA.
spectrin, ankyrin, protein 4.1, and dematin [5]. While much is known about the functions of these proteins, little is known concerning the physiological function of dematin in vivo. In vitro, dematin bundles actin ¢laments in a phosphorylation-dependent manner [6]. While dematin is a substrate of multiple protein kinases, only phosphorylation by the cAMP-dependent protein kinases abolishes its actin-bundling activity [6]. Currently, we are investigating the function of dematin in non-erythroid cells. Dematin transcripts are widely expressed, particularly in the heart, brain, and skeletal muscle [1]. We have also mapped the locus encoding the dematin gene to human chromosome 8p21.1 [2]. In a review of the literature, the chromosomal region 8p21.1 is often deleted in a variety of cancers, including prostate cancer, which suggests the presence of a tumor suppressor gene(s) [7,8]. Here we report the complete structure of the human dematin gene. The availability of dematin's intron^exon organization will be useful for the analysis of possible genetic abnormalities of the dematin
0167-4781 / 98 / $19.00 ß 1998 Elsevier Science B.V. All rights reserved. PII: S 0 1 6 7 - 4 7 8 1 ( 9 8 ) 0 0 0 7 8 - 5
BBAEXP 91146 1-7-98
A.C. Kim et al. / Biochimica et Biophysica Acta 1398 (1998) 382^386
383
Fig. 1. Nucleotide sequence of the intron^exon boundaries of the dematin gene. Nucleotide positions of the intron^exon boundaries correspond to the 52 kDa dematin cDNA (accession no. U28389).
gene in human cancers. We also report the ¢nding of two novel isoforms of dematin in the brain, both lacking exon 2. We used a previously isolated P1 clone to determine the genomic structure of the dematin gene [2]. P1#77 was digested with KpnI and shotgun sub-
cloned into the vector pGEM-4Z. Two of the KpnI subclones contained the entire dematin gene by PCR analysis. Intron^exon boundaries were determined by sequencing the KpnI subclones using primers derived from the dematin cDNA (Fig. 1). Intron sizes were determined by a combination of PCR and DNA
Fig. 2. Genomic organization of the human dematin gene. (A) Full-length erythroid dematin cDNA (52 kDa). (B) Dematin isoforms in erythrocytes and brain. (C) The intron^exon map of the human dematin gene derived from P1 genomic clone #77. Exons 2 and 13 (¢lled black) are alternative coding exons.
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Fig. 3. Expression of dematin mRNA in various tissues. Dematin is primarily expressed as two transcripts of 1.35 and 2.5 kb in length. The 2.5 kb transcript is particularly abundant in the prostate (lane 3), testis (lane 4), and small intestines (lane 6). The lower panel shows the same blot hybridized with L-actin to normalize for loading di¡erences.
sequence analysis. The dematin gene consists of 15 exons interrupted by 14 introns and spans approximately 15 kb (Fig. 2). Exon 13 encodes an alternatively spliced 22 amino acid insertion found in the 52 kDa subunit of erythroid dematin [2]. The PEST sequence, a motif found in signal transduction proteins that undergo rapid turnover in vivo, spans the boundary between exons 4 and 5. A negatively charged cluster of 10 glutamic and aspartic acid residues, a motif commonly found in proteins associated with the cell nucleus, is encoded by exon 8.
We used the BLAST 2.0 program to query the dematin cDNA against the human expressed sequence tags (ESTs) database at the National Center for Biotechnology Information (NCBI) website [9]. Two ESTs (accession nos. H14221 and R87550) matched perfectly with dematin and contained identical gaps in their sequence. Both EST sequences are derived from the 5P end of dematin cDNAs cloned from a human brain cDNA library. Complete DNA sequencing of the cDNAs revealed both to be novel full-length isoforms of dematin. EST R87550 encodes a cDNA that lacks exon 2 while EST H14221 lacks exons 2 and 13 (Fig. 2). The functional signi¢cance of dematin transcripts lacking exon 2 or exons 2 and 13 is presently unknown. Sequence analysis of the 25 amino acids that exon 2 encodes reveals no obvious homologies to any sequences deposited in GenBank. To analyze the expression of dematin in non-erythroid cells, we used the full-length dematin cDNA to probe the Human Multiple Tissue Northern Blot II using standard techniques (Clontech). Each lane is loaded with V2 Wg poly A RNA (Fig. 3). While villin is expressed exclusively in the brush border of intestinal and kidney cells, dematin transcripts are widely expressed, particularly so in the prostate and intestinal tissues. Two major transcripts of V1.35 and 2.5 kb in length hybridized to the cDNA probe. The 2.5 kb message is the predominant transcript in all tissues except in the thymus, and is consistent in size with the reported dematin full-length cDNA [1]. The cloning of dematin revealed it to be the ¢rst protein with sequence homology to the C-terminal headpiece domain of villin [1]. Since then, several proteins have been identi¢ed with similar modules,
Fig. 4. Amino acid sequence alignment of the headpiece domains of human dematin, villin, and limatin. Alignment was performed using the MegAlign software (DNAStar). Villin lacks the 22 amino acid insertion found in dematin and limatin.
BBAEXP 91146 1-7-98
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including limatin and supervillin [3,10]. The dematin headpiece displays 32% amino acid identity with villin and 53% identity with limatin (Fig. 4). Interestingly, limatin is the only headpiece-containing protein to also contain the 22 amino acid insertion found in the 52 kDa dematin headpiece. The dematin headpiece insertion contains the phosphate binding consensus sequence GxGxxGR (P-loop) and has been shown to bind ATP in vitro [11]. Whether limatin similarly binds to nucleotides via the headpiece insertion in currently unknown. However, nucleotide binding seems unlikely since the limatin insertion varies considerably from the P-loop consensus sequence. Previously we, along with other groups, have demonstrated that the headpiece domain binds to actin ¢laments [2,3,12]. However, there is some controversy concerning the physiological function of the headpiece domain in vivo. While transfection studies of mutant villin cDNAs revealed the headpiece to be crucial in the morphogenesis of microvilli, recent evidence from knockout mice suggests that villin plays a minor or redundant role in the development of microvilli in absorptive tissues [12,13]. These studies raise the possibility that in villin null mice, dematin and/or other headpiece-containing proteins may substitute for villin in the reorganization of actin ¢laments in the absorptive epithelia. The short arm of chromosome 8 has been a region of intense scrutiny because of its propensity to undergo deletion in a variety of human cancers [7,8]. Recent evidence has been mounting implicating cytoskeletal proteins as tumor suppressors. Altered expression of the actin-binding proteins tropomyosin, gelsolin, K-actinin, and vinculin suppress the tumorigenesis of epithelial cells [14]. The brain tumor suppressor merlin/schwannomin, a member of the cytoskeletal protein 4.1 superfamily, suppresses the growth of NIH 3T3 cells [15]. The location of the dematin gene at 8p21.1 suggests that dematin may similarly be involved in the regulation of cell proliferation via its ability to organize actin bundles. In fact, we have recently observed the loss of one allele of the dematin gene in a majority of 8p21-linked prostate tumors [16]. Now that the intron^exon structure of the dematin gene has been elucidated, it will be possible to analyze whether the dematin gene is genetically altered in human cancers display-
385
ing loss of heterozygosity of chromosome 8p21.1. It is intriguing to note here that the limatin gene is located at chromosome 10q25, another region frequently deleted in human cancers, including prostate cancer [4]. This work was supported by NIH Grants HL51445 and CA66263 (A.H.C.). A.H.C. is an Established Investigator of the American Heart Association.
References [1] A.P. Rana, P. Ru¡, G.J. Maalouf, D.W. Speicher, A.H. Chishti, Cloning of human erythroid dematin reveals another member of the villin family, Proc. Natl. Acad. Sci. USA 90 (1993) 6651^6655. [2] A.C. Azim, J.H.M. Knoll, A.H. Beggs, A.H. Chishti, Isoform cloning, actin binding, and chromosomal localization of human erythroid dematin, a member of the villin superfamily, J. Biol. Chem. 270 (1995) 17407^17413. [3] D.J. Roof, A. Hayes, M. Adamian, A.H. Chishti, T. Li, Molecular characterization of abLIM, a novel actin-binding and double zinc ¢nger protein, J. Cell Biol. 138 (1997) 575^ 588. [4] A.C. Kim, L. L Peters, J.H.M. Knoll, C. Van Hu¡el, S.L. Ciciotte, P.W. Kleyn, A.H. Chishti, Limatin (LIMAB1), an actin-binding LIM protein, maps to mouse Chromosome 19 and human Chromosome 10q25, a region frequently deleted in human cancers, Genomics 46 (1997) 291^293. [5] D.L. Siegel, D. Branton, Partial puri¢cation and characterization of an actin-binding protein, band 4.9, from human erythrocytes, J. Cell Biol. 100 (1985) 775^785. [6] A.H. Chishti, A. Levin, D. Branton, Abolition of actin-bundling by phosphorylation of human erythrocyte protein 4.9, Nature 334 (1988) 718^721. [7] S.F. Huang, S. Xiao, A.A. Renshaw, K.R. Loughlin, T.J. Hudson, J.A. Fletcher, Fluorescence in situ hybridization evaluation of chromosome deletion patterns in prostate cancer, Am. J. Pathol. 149 (1996) 1565^1573. [8] C.D. Vocke, R.O. Pozzatti, D.G. Bostwick, C.D. Florence, S.B. Jennings, S.E. Strup, P.H. Duray, L.A. Liotta, M.R. Emmert-Buck, W.M. Linehan, Analysis of 99 microdissected prostate carcinomas reveals a high frequency of allelic loss on chromosome 8p12-21, Cancer Res. 56 (1996) 2411^ 2416. [9] S.F. Altschul, T.L. Madden, A.A. Scha¡er, J. Zhang, Z. Zhang, W. Miller, D.J. Lipman, Gapped BLAST and PSIBLAST: a new generation of protein database search programs, Nucleic Acids Res. 25 (1997) 3389^3402. [10] K.N. Pestonjamasp, R.K. Pope, J.D. Wulfkuhle, E.J. Luna, Supervillin (p205): a novel membrane-associated, F-actinbinding protein in the villin/gelsolin superfamily, J. Cell Biol. 139 (1997) 1255^1269.
BBAEXP 91146 1-7-98
386
A.C. Kim et al. / Biochimica et Biophysica Acta 1398 (1998) 382^386
[11] A.C. Azim, S.M. Marfatia, C. Korsgren, E. Dotimas, C.M. Cohen, A.H. Chishti, Human erythrocyte dematin and protein 4.2 (Pallidin) are ATP binding proteins, Biochemistry 35 (1996) 3001^3006. [12] E. Friederich, K. Vancompernolle, C. Huet, M. Goethals, J. Finidori, J. Vandekerckhove, D. Louvard, An actin-binding site containing a conserved motif of charged amino acid residues is essential for the morphogenesis e¡ect of villin, Cell 70 (1992) 81^92. [13] K.I. Pinson, L. Dunbar, L. Samuelson, D.L. Gumucio, Targeted disruption of the mouse villin gene does not impair
the morphogenesis of microvilli, Dev. Dyn. 211 (1998) 109^121. [14] A. Ben-Ze'ev, The cytoskeleton of cancer cells, Biochim. Biophys. Acta 780 (1995) 197^212. [15] M. Lutchman, G.A. Rouleau, The neuro¢bromatosis type 2 gene product schwannomin, suppresses growth of NIH 3T3 cells, Cancer Res. 55 (1995) 2270^2274. [16] A.C. Kim, M. Lutchman, S. Pack, Z.P. Zhuang, A.H. Chishti, Overexpression of dematin alters cell morphology of normal and prostate tumor epithelial cells, Mol. Biol. Cell Suppl. 8 (1997) 276a.
BBAEXP 91146 1-7-98