* 176610

PROFILIN 1; PFN1


HGNC Approved Gene Symbol: PFN1

Cytogenetic location: 17p13.2     Genomic coordinates (GRCh38): 17:4,945,649-4,949,087 (from NCBI)


Gene-Phenotype Relationships
Location Phenotype Phenotype
MIM number		 Inheritance Phenotype
mapping key
17p13.2 Amyotrophic lateral sclerosis 18 614808 3

TEXT

▼ Description

Profilin-1 is a 140-amino acid protein and major growth regulator of filamentous (F)-actin through its binding of monomeric (G)-actin (Mockrin and Korn, 1980).


▼ Cloning and Expression

Profilin is a ubiquitous 12- to 15-kD protein which inhibits the polymerization of actin. It is thought to function by complexing with unpolymerized actin in vivo. Dissociation of the profilin-actin complex is caused by binding of phosphatidylinositol 4,5-bisphosphate to profilin. Ampe et al. (1988) described the amino acid sequence of human platelet profilin and found it to have 95% homology to the sequence of calf spleen profilin.


▼ Gene Function

Goldschmidt-Clermont and Janmey (1991) reviewed the function of profilin, partly on the basis of work in the S. cerevisiae, homolog, PFY (Vojtek et al., 1991). Theriot and Mitchison (1993) reviewed the multiple functions of profilin.


▼ Mapping

By Southern blot analysis of somatic cell hybrid DNA, Kwiatkowski and Bruns (1988) found that at least 4 dispersed genetic loci in the human genome hybridize with the profilin cDNA as well as the untranslated region fragments, suggesting that several of these loci represent pseudogenes of recent evolutionary origin. Chromosomes 1, 6, 13, and 17 were implicated. In addition, 5-prime and 3-prime untranslated regions were found to be conserved between humans and rodents, implying a functional role for these regions of the profilin gene. Kwiatkowski et al. (1990) localized the functional profilin gene to 17p13 by analysis of somatic cell hybrids and by in situ hybridization. By study of patients with deletions and by use of somatic cell hybrids containing a deleted chromosome 17, Kwiatkowski et al. (1990) sublocalized the PFN1 gene to 17p13.3. This is the same region as that deleted in the Miller-Dieker syndrome (MDLS; 247200). They found that the gene indeed was deleted in some patients with MDLS but that other patients had smaller deletions not affecting the profilin locus. Thus, a single allelic deletion of the profilin locus may contribute to the clinical phenotype of MDLS in some patients but does not play a major role in the essential phenotype.

In the mouse, Klingenspor et al. (1997) mapped the Pfn1 gene to chromosome 11 and a profilin-1 related sequence to chromosome 15.


▼ Molecular Genetics

Wu et al. (2012) identified 4 different mutations in the PFN1 gene in 7 of 274 familial ALS cases. One mutation (E117G; 176610.0004), was found in 3 of 1,090 ALS cases and 3 of 7,560 controls. Wu et al. (2012) performed in vitro assays and showed that mutant PFN1 produces ubiquitinated, insoluble aggregates in transfected cells. In many cases the aggregates contained the ALS-associated protein TDP43 (605078). The E117G mutation (176610.0004) displayed a pattern more similar to wildtype PFN1, with most of the expressed protein in the soluble fraction. Wu et al. (2012) found that mutant PFN1 inhibited axon outgrowth. Wu et al. (2012) concluded that mutations in the PFN1 gene account for approximately 1 to 2% of familial ALS and suggested that disruption of cytoskeletal pathways contribute importantly to ALS pathogenicity.


▼ Animal Model

To examine the function of profilin-1 in vivo, Witke et al. (2001) generated Pfn1 knockout mice. Homozygotes were not viable; they died as early as the 2-cell stage, and no homozygous knockout blastocysts were detectable. Adult heterozygotes showed a 50% reduction in profilin-1 expression with no apparent impairment of cell function. However, heterozygous embryos had reduced survival during embryogenesis compared with wildtype. Although weakly expressed in early embryos, profilin-2 (176590) could not compensate for lack of profilin-1. Their results indicated that mouse profilin-1 is an essential protein that has dosage-dependent effects on cell division and survival during embryogenesis.

Yarovinsky et al. (2005) identified a profilin-like molecule from the protozoan parasite Toxoplasma gondii that generates a potent IL12 (see 161560) response in murine dendritic cells that is dependent on MyD88 (602170). T. gondii profilin activates dendritic cells through TLR11 (606270) and is the first chemically defined ligand for this toll-like receptor. Moreover, TLR11 is required in vivo for parasite-induced IL12 production and optimal resistance to infection, thereby establishing a role for the receptor in host recognition of protozoan pathogens.


▼ ALLELIC VARIANTS ( 4 Selected Examples):

.0001  AMYOTROPHIC LATERAL SCLEROSIS 18

PFN1, CYS71GLY   
dbSNP:rs387907264 RCV000030694

In 3 families segregating autosomal dominant ALS18 (614808), Wu et al. (2012) identified a T-to-G transversion at nucleotide 347 of the PFN1 gene, resulting in a cysteine-to-glycine substitution at codon 71 (C71G). In family 1, all 4 affected members for which DNA was available possessed the C71G variant. A single obligate carrier of the C71G variant did not develop disease, but she died before the average age of onset of this family (50.0 +/- 6.6 years). All unaffected family members displayed the wildtype genotype. This mutation was subsequently identified in 2 other families. In family 3, the mutation was detected in 3 other affected family members. A single unaffected member of the family whose age was in the mid-forties was found to be a mutation carrier. The average age of onset in this family was 41.1 +/- 4.3 years for those definitively affected. Haplotype analysis suggested that the C71G variant derives from a single ancestral mutation. This mutation was not observed among 7,560 control samples (15,120 alleles).


.0002  AMYOTROPHIC LATERAL SCLEROSIS 18

PFN1, MET114THR   
dbSNP:rs387907265 RCV000030695

In 2 families (families 2 and 4) segregating autosomal dominant ALS18 (614808), Wu et al. (2012) identified a T-to-C transition at nucleotide 477 of the PFN1 gene, resulting in a methionine-to-threonine substitution at codon 114 (M114T). In family 2, all 8 affected members for whom DNA was available carried the mutation, and on the basis of genotype in spouse and progeny, Wu et al. (2012) were able to confirm that a ninth affected family member also carried the mutation. Of 7 unaffected members, 5 did not carry the M114T variant. One unaffected mutation carrier was aged in the mid-forties and a second obligate carrier was asymptomatic into the seventies, suggesting high but incomplete penetrance of this mutation. The average age of onset for family 2 was 41.9 +/- 5.3 years and for family 4 it was 52.0 +/- 13.1 years. This mutation was not observed among 7,560 control samples (15,120 alleles).


.0003  AMYOTROPHIC LATERAL SCLEROSIS 18

PFN1, GLY118VAL   
dbSNP:rs387907266 RCV000030696

In an individual from a family with a parent-child segregating autosomal dominant ALS18 (614808), Wu et al. (2012) identified a G-to-T transversion at nucleotide 489 of the PFN1 gene, resulting in a glycine-to-valine substitution at codon 118 (G118V). This mutation was not observed among 7,560 control samples (15,120 alleles). The glycine at this position is invariant down to zebrafish.


.0004  AMYOTROPHIC LATERAL SCLEROSIS 18

In one family with autosomal dominant ALS (ALS18; 614808) with incomplete penetrance, Wu et al. (2012) identified a 2-basepair substitution, AA to GT, at nucleotides 46 and 47 of the PFN1 gene, resulting in a glutamic acid-to-glycine substitution at codon 117 (E117G). This mutation was subsequently identified in 2 sporadic ALS patients among 816 sequenced. Overall, this mutation was identified in 3 of 1,090 ALS cases and 3 of 7,560 control samples (2.75 x 10(-3) vs 3.97 x 10(-4); P = 0.030, two-tailed Fisher's exact test). Wu et al. (2012) suggested that, while it could be argued that the E117G variant is nonpathogenic, more likely it is less pathogenic than the other 3 mutations (C71G; 176610.0001, M114T; 176610.0002, and G118V; 176610.0003) identified in the PFN1 gene to that time. While Western blot analysis of the soluble and insoluble fractions of cells transfected with wildtype and mutant PFN1 proteins detected a considerable portion of the C71G, M114T, and G118V mutant proteins in the insoluble fraction, the E117G mutation displayed a pattern more similar to wildtype PFN1, with most of the expressed protein in the soluble fraction. The glu at position 117 is invariant down to zebrafish.


▼ REFERENCES

  1. Ampe, C., Markey, F., Lindberg, U., Vandekerckhove, J. The primary structure of human platelet profilin: reinvestigation of the calf spleen profilin sequence. FEBS Lett. 228: 17-21, 1988. [PubMed: 3342873, related citations] [Full Text]

  2. Goldschmidt-Clermont, P. J., Janmey, P. A. Profilin, a weak CAP for actin and RAS. Cell 66: 419-421, 1991. [PubMed: 1651167, related citations] [Full Text]

  3. Klingenspor, M., Bodnar, J., Xia, Y.-R., Welch, C., Lusis, A. J., Reue, K. Localization of profilin-1 (Pfn1) and a related sequence (Pfn1-rs) to mouse chromosomes 11 and 15 respectively. Mammalian Genome 8: 539-541, 1997. [PubMed: 9196007, related citations]

  4. Kwiatkowski, D. J., Aklog, L., Ledbetter, D. H., Morton, C. C. Identification of the functional profilin gene, its localization to chromosome subband 17p13.3, and demonstration of its deletion in some patients with Miller-Dieker syndrome. Am. J. Hum. Genet. 46: 559-567, 1990. [PubMed: 1968707, related citations]

  5. Kwiatkowski, D. J., Bruns, G. A. P. Human profilin: molecular cloning, sequence comparison, and chromosomal analysis. J. Biol. Chem. 263: 5910-5915, 1988. [PubMed: 3356709, related citations] [Full Text]

  6. Mockrin, S. C., Korn, E. D. Acanthamoeba profilin interacts with G-actin to increase the rate of exchange of actin-bound adenosine 5-prime-triphosphate. Biochemistry 19: 5359-5362, 1980. [PubMed: 6893804, related citations]

  7. Theriot, J. A., Mitchison, T. J. The three faces of profilin. Cell 75: 835-838, 1993. [PubMed: 8252619, related citations] [Full Text]

  8. Vojtek, A., Haarer, B., Field, J., Gerst, J., Pollard, T. D., Brown, S., Wigler, M. Evidence for a functional link between profilin and CAP in the yeast S. cerevisiae. Cell 66: 497-505, 1991. [PubMed: 1868547, related citations] [Full Text]

  9. Witke, W., Sutherland, J. D., Sharpe, A., Arai, M., Kwiatkowski, D. J. Profilin I is essential for cell survival and cell division in early mouse development. Proc. Nat. Acad. Sci. 98: 3832-3836, 2001. [PubMed: 11274401, images, related citations] [Full Text]

  10. Wu, C.-H., Fallini, C., Ticozzi, N., Keagle, P. J., Sapp, P. C., Piotrowska, K., Lowe, P., Koppers, M., McKenna-Yasek, D., Baron, D. M., Kost, J. E., Gonzalez-Perez, P., and 26 others. Mutations in the profilin 1 gene cause familial amyotrophic lateral sclerosis. Nature 488: 499-503, 2012. [PubMed: 22801503, images, related citations] [Full Text]

  11. Yarovinsky, F., Zhang, D., Andersen, J. F., Bannenberg, G. L., Serhan, C. N., Hayden, M. S., Hieny, S., Sutterwala, F. S., Flavell, R. A., Ghosh, S., Sher, A. TLR11 activation of dendritic cells by a protozoan profilin-like protein. Science 308: 1626-1629, 2005. [PubMed: 15860593, related citations] [Full Text]


Contributors:
Ada Hamosh - updated : 9/5/2012
Ada Hamosh - updated : 2/6/2006
Victor A. McKusick - updated : 4/17/2001
Victor A. McKusick - updated : 8/18/1997
Creation Date:
Victor A. McKusick : 4/23/1988
Edit History:
alopez : 09/06/2012
terry : 9/5/2012
alopez : 2/6/2006
mcapotos : 5/8/2001
mcapotos : 4/24/2001
terry : 4/17/2001
dkim : 7/24/1998
terry : 8/18/1997
mimadm : 2/25/1995
carol : 4/28/1994
warfield : 3/31/1994
carol : 12/16/1993
carol : 10/26/1993
supermim : 3/16/1992

* 176610

PROFILIN 1; PFN1


HGNC Approved Gene Symbol: PFN1

Cytogenetic location: 17p13.2     Genomic coordinates (GRCh38): 17:4,945,649-4,949,087 (from NCBI)


Gene-Phenotype Relationships

Location Phenotype Phenotype
MIM number		 Inheritance Phenotype
mapping key
17p13.2 Amyotrophic lateral sclerosis 18 614808 3

TEXT

Description

Profilin-1 is a 140-amino acid protein and major growth regulator of filamentous (F)-actin through its binding of monomeric (G)-actin (Mockrin and Korn, 1980).


Cloning and Expression

Profilin is a ubiquitous 12- to 15-kD protein which inhibits the polymerization of actin. It is thought to function by complexing with unpolymerized actin in vivo. Dissociation of the profilin-actin complex is caused by binding of phosphatidylinositol 4,5-bisphosphate to profilin. Ampe et al. (1988) described the amino acid sequence of human platelet profilin and found it to have 95% homology to the sequence of calf spleen profilin.


Gene Function

Goldschmidt-Clermont and Janmey (1991) reviewed the function of profilin, partly on the basis of work in the S. cerevisiae, homolog, PFY (Vojtek et al., 1991). Theriot and Mitchison (1993) reviewed the multiple functions of profilin.


Mapping

By Southern blot analysis of somatic cell hybrid DNA, Kwiatkowski and Bruns (1988) found that at least 4 dispersed genetic loci in the human genome hybridize with the profilin cDNA as well as the untranslated region fragments, suggesting that several of these loci represent pseudogenes of recent evolutionary origin. Chromosomes 1, 6, 13, and 17 were implicated. In addition, 5-prime and 3-prime untranslated regions were found to be conserved between humans and rodents, implying a functional role for these regions of the profilin gene. Kwiatkowski et al. (1990) localized the functional profilin gene to 17p13 by analysis of somatic cell hybrids and by in situ hybridization. By study of patients with deletions and by use of somatic cell hybrids containing a deleted chromosome 17, Kwiatkowski et al. (1990) sublocalized the PFN1 gene to 17p13.3. This is the same region as that deleted in the Miller-Dieker syndrome (MDLS; 247200). They found that the gene indeed was deleted in some patients with MDLS but that other patients had smaller deletions not affecting the profilin locus. Thus, a single allelic deletion of the profilin locus may contribute to the clinical phenotype of MDLS in some patients but does not play a major role in the essential phenotype.

In the mouse, Klingenspor et al. (1997) mapped the Pfn1 gene to chromosome 11 and a profilin-1 related sequence to chromosome 15.


Molecular Genetics

Wu et al. (2012) identified 4 different mutations in the PFN1 gene in 7 of 274 familial ALS cases. One mutation (E117G; 176610.0004), was found in 3 of 1,090 ALS cases and 3 of 7,560 controls. Wu et al. (2012) performed in vitro assays and showed that mutant PFN1 produces ubiquitinated, insoluble aggregates in transfected cells. In many cases the aggregates contained the ALS-associated protein TDP43 (605078). The E117G mutation (176610.0004) displayed a pattern more similar to wildtype PFN1, with most of the expressed protein in the soluble fraction. Wu et al. (2012) found that mutant PFN1 inhibited axon outgrowth. Wu et al. (2012) concluded that mutations in the PFN1 gene account for approximately 1 to 2% of familial ALS and suggested that disruption of cytoskeletal pathways contribute importantly to ALS pathogenicity.


Animal Model

To examine the function of profilin-1 in vivo, Witke et al. (2001) generated Pfn1 knockout mice. Homozygotes were not viable; they died as early as the 2-cell stage, and no homozygous knockout blastocysts were detectable. Adult heterozygotes showed a 50% reduction in profilin-1 expression with no apparent impairment of cell function. However, heterozygous embryos had reduced survival during embryogenesis compared with wildtype. Although weakly expressed in early embryos, profilin-2 (176590) could not compensate for lack of profilin-1. Their results indicated that mouse profilin-1 is an essential protein that has dosage-dependent effects on cell division and survival during embryogenesis.

Yarovinsky et al. (2005) identified a profilin-like molecule from the protozoan parasite Toxoplasma gondii that generates a potent IL12 (see 161560) response in murine dendritic cells that is dependent on MyD88 (602170). T. gondii profilin activates dendritic cells through TLR11 (606270) and is the first chemically defined ligand for this toll-like receptor. Moreover, TLR11 is required in vivo for parasite-induced IL12 production and optimal resistance to infection, thereby establishing a role for the receptor in host recognition of protozoan pathogens.


ALLELIC VARIANTS 4 Selected Examples):

.0001  AMYOTROPHIC LATERAL SCLEROSIS 18

PFN1, CYS71GLY    rs387907264

In 3 families segregating autosomal dominant ALS18 (614808), Wu et al. (2012) identified a T-to-G transversion at nucleotide 347 of the PFN1 gene, resulting in a cysteine-to-glycine substitution at codon 71 (C71G). In family 1, all 4 affected members for which DNA was available possessed the C71G variant. A single obligate carrier of the C71G variant did not develop disease, but she died before the average age of onset of this family (50.0 +/- 6.6 years). All unaffected family members displayed the wildtype genotype. This mutation was subsequently identified in 2 other families. In family 3, the mutation was detected in 3 other affected family members. A single unaffected member of the family whose age was in the mid-forties was found to be a mutation carrier. The average age of onset in this family was 41.1 +/- 4.3 years for those definitively affected. Haplotype analysis suggested that the C71G variant derives from a single ancestral mutation. This mutation was not observed among 7,560 control samples (15,120 alleles).


.0002  AMYOTROPHIC LATERAL SCLEROSIS 18

PFN1, MET114THR    rs387907265

In 2 families (families 2 and 4) segregating autosomal dominant ALS18 (614808), Wu et al. (2012) identified a T-to-C transition at nucleotide 477 of the PFN1 gene, resulting in a methionine-to-threonine substitution at codon 114 (M114T). In family 2, all 8 affected members for whom DNA was available carried the mutation, and on the basis of genotype in spouse and progeny, Wu et al. (2012) were able to confirm that a ninth affected family member also carried the mutation. Of 7 unaffected members, 5 did not carry the M114T variant. One unaffected mutation carrier was aged in the mid-forties and a second obligate carrier was asymptomatic into the seventies, suggesting high but incomplete penetrance of this mutation. The average age of onset for family 2 was 41.9 +/- 5.3 years and for family 4 it was 52.0 +/- 13.1 years. This mutation was not observed among 7,560 control samples (15,120 alleles).


.0003  AMYOTROPHIC LATERAL SCLEROSIS 18

PFN1, GLY118VAL    rs387907266

In an individual from a family with a parent-child segregating autosomal dominant ALS18 (614808), Wu et al. (2012) identified a G-to-T transversion at nucleotide 489 of the PFN1 gene, resulting in a glycine-to-valine substitution at codon 118 (G118V). This mutation was not observed among 7,560 control samples (15,120 alleles). The glycine at this position is invariant down to zebrafish.


.0004  AMYOTROPHIC LATERAL SCLEROSIS 18

PFN1, GLU117GLY    rs140547520

In one family with autosomal dominant ALS (ALS18; 614808) with incomplete penetrance, Wu et al. (2012) identified a 2-basepair substitution, AA to GT, at nucleotides 46 and 47 of the PFN1 gene, resulting in a glutamic acid-to-glycine substitution at codon 117 (E117G). This mutation was subsequently identified in 2 sporadic ALS patients among 816 sequenced. Overall, this mutation was identified in 3 of 1,090 ALS cases and 3 of 7,560 control samples (2.75 x 10(-3) vs 3.97 x 10(-4); P = 0.030, two-tailed Fisher's exact test). Wu et al. (2012) suggested that, while it could be argued that the E117G variant is nonpathogenic, more likely it is less pathogenic than the other 3 mutations (C71G; 176610.0001, M114T; 176610.0002, and G118V; 176610.0003) identified in the PFN1 gene to that time. While Western blot analysis of the soluble and insoluble fractions of cells transfected with wildtype and mutant PFN1 proteins detected a considerable portion of the C71G, M114T, and G118V mutant proteins in the insoluble fraction, the E117G mutation displayed a pattern more similar to wildtype PFN1, with most of the expressed protein in the soluble fraction. The glu at position 117 is invariant down to zebrafish.


REFERENCES

  1. Ampe, C., Markey, F., Lindberg, U., Vandekerckhove, J. The primary structure of human platelet profilin: reinvestigation of the calf spleen profilin sequence. FEBS Lett. 228: 17-21, 1988. [PubMed: 3342873] [Full Text: https://linkinghub.elsevier.com/retrieve/pii/0014-5793(88)80575-1]

  2. Goldschmidt-Clermont, P. J., Janmey, P. A. Profilin, a weak CAP for actin and RAS. Cell 66: 419-421, 1991. [PubMed: 1651167] [Full Text: https://linkinghub.elsevier.com/retrieve/pii/0092-8674(81)90002-7]

  3. Klingenspor, M., Bodnar, J., Xia, Y.-R., Welch, C., Lusis, A. J., Reue, K. Localization of profilin-1 (Pfn1) and a related sequence (Pfn1-rs) to mouse chromosomes 11 and 15 respectively. Mammalian Genome 8: 539-541, 1997. [PubMed: 9196007]

  4. Kwiatkowski, D. J., Aklog, L., Ledbetter, D. H., Morton, C. C. Identification of the functional profilin gene, its localization to chromosome subband 17p13.3, and demonstration of its deletion in some patients with Miller-Dieker syndrome. Am. J. Hum. Genet. 46: 559-567, 1990. [PubMed: 1968707]

  5. Kwiatkowski, D. J., Bruns, G. A. P. Human profilin: molecular cloning, sequence comparison, and chromosomal analysis. J. Biol. Chem. 263: 5910-5915, 1988. [PubMed: 3356709] [Full Text: http://www.jbc.org/cgi/pmidlookup?view=long&pmid=3356709]

  6. Mockrin, S. C., Korn, E. D. Acanthamoeba profilin interacts with G-actin to increase the rate of exchange of actin-bound adenosine 5-prime-triphosphate. Biochemistry 19: 5359-5362, 1980. [PubMed: 6893804]

  7. Theriot, J. A., Mitchison, T. J. The three faces of profilin. Cell 75: 835-838, 1993. [PubMed: 8252619] [Full Text: https://linkinghub.elsevier.com/retrieve/pii/0092-8674(93)90527-W]

  8. Vojtek, A., Haarer, B., Field, J., Gerst, J., Pollard, T. D., Brown, S., Wigler, M. Evidence for a functional link between profilin and CAP in the yeast S. cerevisiae. Cell 66: 497-505, 1991. [PubMed: 1868547] [Full Text: https://linkinghub.elsevier.com/retrieve/pii/0092-8674(81)90013-1]

  9. Witke, W., Sutherland, J. D., Sharpe, A., Arai, M., Kwiatkowski, D. J. Profilin I is essential for cell survival and cell division in early mouse development. Proc. Nat. Acad. Sci. 98: 3832-3836, 2001. [PubMed: 11274401] [Full Text: http://www.pnas.org/cgi/pmidlookup?view=long&pmid=11274401]

  10. Wu, C.-H., Fallini, C., Ticozzi, N., Keagle, P. J., Sapp, P. C., Piotrowska, K., Lowe, P., Koppers, M., McKenna-Yasek, D., Baron, D. M., Kost, J. E., Gonzalez-Perez, P., and 26 others. Mutations in the profilin 1 gene cause familial amyotrophic lateral sclerosis. Nature 488: 499-503, 2012. [PubMed: 22801503] [Full Text: https://dx.doi.org/10.1038/nature11280]

  11. Yarovinsky, F., Zhang, D., Andersen, J. F., Bannenberg, G. L., Serhan, C. N., Hayden, M. S., Hieny, S., Sutterwala, F. S., Flavell, R. A., Ghosh, S., Sher, A. TLR11 activation of dendritic cells by a protozoan profilin-like protein. Science 308: 1626-1629, 2005. [PubMed: 15860593] [Full Text: http://www.sciencemag.org/cgi/pmidlookup?view=long&pmid=15860593]


Contributors:
Ada Hamosh - updated : 9/5/2012
Ada Hamosh - updated : 2/6/2006
Victor A. McKusick - updated : 4/17/2001
Victor A. McKusick - updated : 8/18/1997
Creation Date:
Victor A. McKusick : 4/23/1988
Edit History:
alopez : 09/06/2012
terry : 9/5/2012
alopez : 2/6/2006
mcapotos : 5/8/2001
mcapotos : 4/24/2001
terry : 4/17/2001
dkim : 7/24/1998
terry : 8/18/1997
mimadm : 2/25/1995
carol : 4/28/1994
warfield : 3/31/1994
carol : 12/16/1993
carol : 10/26/1993
supermim : 3/16/1992