Genetic marker for spondyloepimetaphyseal dysplasia

A nucleic acid construct comprising a nucleic acid segment comprising:

• (A) a nucleotide sequence of (SEQ. ID. NO.:1), a nucleotide sequence complementary thereto, a degenerate coding sequence thereof, or a gene-specific fragment of any of these;
• or
• (B) a nucleic acid segment encoding a human PAPSS2 protein having an amino acid sequence of (SEQ. ID. NO.:7).

An isolated polynucleotide comprising a nucleic acid segment encoding a murine PAPS synthetase, said nucleic acid segment having a nucleotide sequence of (SEQ. ID. NO.:2), a nucleotide sequence complementary thereto, a degenerate coding sequence thereof, or a gene-specific fragment of any of these.

A nucleic acid construct, comprising a nucleic acid segment comprising:

• (A) a nucleotide sequence of (SEQ. ID. NO.:2), a nucleotide sequence complementary thereto, a degenerate coding sequence thereof, or a gene-specific fragment of any of these;
• or
• (B) a nucleic acid segment encoding a human PAPSS2 protein having an amino acid sequence of(SEQ. ID. NO.:8).

An isolated PAPS synthetase protein comprising a polypeptide having an amino acid sequence of (SEQ. ID. NO.:7), (SEQ. ID. NO.:8), or an antibody binding fragment of either of these at least 6 amino acids long.

A PAPSS2 fusion protein, comprising:

• a first PAPSS2 polypeptide segment comprising an amino acid sequence of (SEQ. ID. NO.:7) or a gene-specific antibody binding fragment thereof at least 6 amino acids long;
• and
• a second predetermined polypeptide segment.

A Papss2 fusion protein comprising:

• a first Papss2 polypeptide segment comprising an amino acid sequence of (SEQ. ID. NO.:8) or a gene-specific antibody binding fragment thereof at least 6 amino acids long;
• and
• a second predetermined polypeptide segment.

A method of diagnosing spondyloepimetaphyseal dysplasia in a human subject, comprising:

• a) amplifying a nucleic acid segment from a sample of a bodily substance containing human nucleic acid, said sample being derived from a human subject having at least one symptom of spondyloepimetaphyseal dysplasia, said nucleic acid segment defining a sequence from human chromosomal region 10q23-24, between microsatellite markers D10S1143 and D10S2470, to produce amplification products;
• and
• b) analyzing the amplification products for the presence of homozygosity for a variant allele of a PAPSS2 gene, the presence of homozygosity for the variant allele of the gene corroborating a diagnosis of spondyloepimetaphyseal dysplasia in the human subject.

A method of diagnosing spondyloepimetaphyseal dysplasia Pakistani-type in a human subject, comprising:

• a) amplifying a nucleic acid segment from a sample of a bodily substance containing human nucleic acid, said sample being derived from a human subject having at least one symptom of spondyloepimetaphyseal dysplasia Pakistani-type, said nucleic acid segment defining a sequence from human chromosomal region 10q23-24, between microsatellite markers D10S1143 and D10S2470, to produce amplification products;
• and
• b) analyzing the amplification products for the presence of homozygosity for a variant allele of a gene encoding a PAPS synthetase, said variant allele defining a stop codon instead of a serine codon corresponding to amino acid residue 475 of SEQ. ID. NO.:7, the presence of homozygosity for the variant allele corroborating a diagnosis of spondyloepimetaphyseal dysplasia Pakistani-type in the human subject.

A method of identifying a human carrier of an heritable allele associated with spondyloepimetaphyseal dysplasia, comprising:

• a) amplifying a nucleic acid segment from a sample of a bodily substance containing human nucleic acid, said sample being derived from a human subject without a symptom of spondyloepimetaphyseal dysplasia, said nucleic acid segment defining a sequence from human chromosomal region 10q23-24, between microsatellite markers D10S1143 and D10S2470, to produce amplification products;
• and
• b) analyzing the amplification products for the presence of a variant allele of a gene encoding a PAPS synthetase, the presence of the variant allele of the gene identifying the human subject as a carrier of an heritable allele associated with spondyloepimetaphyseal dysplasia.

A method of identifying a human carrier of an heritable allele associated with spondyloepimetaphyseal dysplasia Pakistani-type, comprising:

• a) amplifying a nucleic acid segment from a sample of a bodily substance containing human nucleic acid, said sample being derived from a human subject without a symptom of spondyloepimetaphyseal dysplasia Pakistani-type, said nucleic acid segment defining a sequence from human chromosomal region 10q23-24, between microsatellite markers D10S1143 and D10S2470, to produce amplification products;
• and
• b) analyzing the amplification products for the presence of homozygosity for a variant allele of a gene encoding a PAPS synthetase, said variant allele defining a stop codon instead of a serine codon corresponding to amino acid residue 475 of SEQ. ID. NO.:7, the presence of the variant allele of the gene identifying the human subject as a carrier of an heritable allele associated with spondyloepimetaphyseal dysplasia Pakistani-type.

A kit for genetically modifying a vertebrate cell, comprising a polynucleotide that comprises a PAPSS2 sequence having (SEQ. ID. NOS.:1), a sequence complementary thereto, or a degenerate coding sequence thereof, or a gene-specific fragment of any of these;

• and
• instructions for using the polynucleotide for genetically modifying a vertebrate cell.

A kit for genetically modifying a vertebrate cell, comprising a polynucleotide that comprises a Papss2 sequence having (SEQ. ID. NOS.:2), a sequence complementary thereto, or a degenerate coding sequence thereof, or a gene-specific fragment of any of these;

• and
• instructions for using the polynucleotide for genetically modifying a vertebrate cell.

An isolated antibody or antibody fragment, comprising an antibody or antibody fragment that selectively binds a PAPS synthetase protein that has an amino acid sequence of (SEQ. ID. NO.:7) or (SEQ.ID. NO.8), or that selectively binds an antibody binding fragment of either of these at least 6 amino acids long.

A protein therapy method for treating a human subject having an osteoarthritic disorder, comprising:

• exposing a cell of a tissue of a human subject having an osteoarthritic disorder that is caused or aggravated by deficient enzymatic sulfation activity to a fusion protein comprising a first PAPSS2 polypeptide segment that comprises an amino acid sequence of (SEQ. ID. NO.:7), or an enzymatically active fragment thereof, and a second polypeptide segment capable of infiltrating the cell, whereby the fusion protein is taken up by the cell and the PAPSS2 polypeptide segment is enzymatically active therein.

A kit for the treatment of osteoarthritic disorders caused or aggravated by deficient enzymatic sulfation activity, comprising:

• a fusion protein comprising a first PAPSS2 polypeptide segment that comprises an amino acid sequence of (SEQ. ID. NO.:7), or an enzymatically active fragment thereof, and a second polypeptide segment capable of infiltrating the cell;
• and
• instructions for using the fusion protein for treating osteoarthritic disorder(s) caused or aggravated by deficient enzymatic sulfation activity.