CIF130 inhibits cell cycle progression

Abstract

A human gene termed CIF130 and its expression products can alter the spatial or temporal patterns of mitosis or cell cycle progression of a human cell. Methods of treating disorders involving alterations in the regulation of mitosis or cell cycle progression utilize the gene and its expression product. Genes whose expression is dependent upon CIF130 expression can be identified.

1 . An isolated and purified human CIF130 protein comprising an amino acid sequence which is at least 85% identical to the amino acid sequence shown in SEQ ID NO: 2, wherein percent identity is determined using a Smith-Waterman homology search algorithm using an affine gap search with a gap open penalty of 12 and a gap extension penalty of 1.  
     
     
         2 . The isolated human CIF130 protein of  claim 1  which has the amino acid sequence shown in SEQ ID NO: 2.  
     
     
         3 . The protein of  claim 1  comprising at least 17 contiguous amino acids as shown in SEQ ID NO: 2.  
     
     
         4 . A CIF130 fusion protein comprising a first protein segment and a second protein segment fused together by means of a peptide bond, wherein the first protein segment consists of the polypeptide of  claim 3 .  
     
     
         5 . A preparation of antibodies which specifically binds to the protein of  claim 2 .  
     
     
         6 . A cDNA molecule which enclodes the protein of  claim 1 .  
     
     
         7 . The cDNA molecule of  claim 6  which encodes at least 17 contiguous amino acids of SEQ ID NO: 2.  
     
     
         8 . The cDNA molecule of  claim 7  which encodes SEQ ID NO: 2.  
     
     
         9 . The cDNA molecule of  claim 8  which comprises SEQ ID NO: 1.  
     
     
         10 . The cDNA molecule of  claim 6  comprising at least 12 contiguous nucleotides of SEQ ID NO: 1.  
     
     
         11 . A cDNA molecule which is at least 85% identical to the nucleotide sequence shown in SEQ ID NO: 1, wherein percent identity is determined using a Smith-Waterman homology search algorithm as implemented in a MPSRCH program using an affine gap search with a gap open penalty of 12 and a gap extension penalty of 1.  
     
     
         12 . An isolated and purified subgenomic polynucleotide comprising a nucleotide sequence which hybridizes to SEQ ID NO: 1 after washing with 0.2× SSC at 65° C., wherein the nucleotide sequence encodes a CIF130 protein having the amino acid sequence of SEQ ID NO: 2.  
     
     
         13 . A construct comprising: 
 a promoter; and    a polynucleotide segment encoding at least 17 contiguous amino acids of a human CIF130 protein as shown in SEQ ID NO: 2, wherein the polynucleotide segment is located downstream from the promoter, wherein transcription of the polynucleotide segment initiates at the promoter.    
     
     
         14 . A host cell comprising the construct of  claim 13 .  
     
     
         15 . A homologously recombinant cell having incorporated therein a new transcription initiation unit, wherein the new transcription initiation unit comprises: 
 (a) an exogenous regulatory sequence;    (b) an exogenous exon; and    (c) a splice donor site, wherein the transcription initiation unit is located upstream of a coding sequence of a CIF130 gene as shown in SEQ ID NO: 1,    wherein the exogenous regulatory sequence directs transcription of the coding sequence of the CIF130 gene.    
     
     
         16 . The homologously recombinant cell of  claim 15  wherein the exogenous regulatory sequence directs enhanced transcription of the coding sequence of the CIF130 gene.  
     
     
         17 . A method to aid in the diagnosis or prognosis of neoplasia in a human, comprising the step of: 
 comparing expression of a first CIF130 gene in a first tissue of a human suspected of being neoplastic with expression of a second CIF130 gene in a second tissue of a human which is normal, wherein increased expression of the first CIF130 gene relative to expression of the second CIF130 gene indicates neoplasia in the first tissue.    
     
     
         18 . A method to aid in the diagnosis or prognosis of neoplasia in a human, comprising the step of: 
 comparing a first human CIF130 gene, mRNA, or protein in a first tissue suspected of being neoplastic with a second human CIF130 gene, mRNA, or protein in a second tissue which is normal, wherein the second CIF130 gene has a coding sequence as shown in SEQ ID NO: 1, wherein a difference between the first and second CIF130 genes, mRNAs, or proteins indicates neoplasia in the first tissue.    
     
     
         19 . A method to aid in detecting a genetic predisposition to neoplasia in a human, comprising: 
 comparing a CIF130 gene, mRNA, or protein in a fetal tissue of a human with a wild-type human CIF130 gene, mRNA, or protein, wherein the wild-type human CIF130 gene has a coding sequence as shown in SEQ ID NO: 1, wherein a difference between the CIF130 gene, mRNA, or protein in the fetal tissue of the human and the wild-type human CIF130 gene, mRNA, or protein indicates a genetic predisposition to neoplasia in the human.    
     
     
         20 . A method of screening test compounds for the ability to interfere with the binding of a CIF130 protein to a CIF150/hTAF II 150 protein, comprising the steps of: 
 (a) contacting a test compound with at least a CIF150/hTAF II 150-binding domain of a CIF130 protein as shown in SEQ ID NO: 2 and at least a CIF130-binding domain of a CIF150/hTAF II 150 protein as shown in SEQ ID NO: 4, wherein the CIF150/hTAF II 150-binding domain binds to the CIF130-binding domain in the absence of the test compound; and    (b) determining the amount of the CIF150/hTAF II 150-binding domain which is bound or unbound to the CIF130-binding domain or determining the amount of the CIF130-binding domain which is bound or unbound to the CIF150/hTAF II 150-binding domain in the presence of the test compound, wherein a test compound which decreases the amount of bound CIF150/hTAF II 150- or CIF130-binding domains or which increases the amount of unbound CIF150/hTAF II 150- and CIF130-binding domains is a potential inducer of mitosis or cell cycle progression.    
     
     
         21 . A method of screening test compounds for the ability to interfere with the binding of a CIF130 protein to a CIF150/hTAF II 150 protein, comprising the steps of: 
 (a) contacting a cell with a test compound, wherein the cell comprises: 
 i) a first fusion protein comprising (1) a CIF150/hTAF II 150-binding domain of a CIF130 protein as shown in SEQ ID NO: 2 and (2) either a DNA binding domain or a transcriptional activating domain;  
 ii) a second fusion protein comprising a CIF130-binding domain of a CIF150/hTAF II 150 protein as shown in SEQ ID NO: 4, wherein the CIF130-binding domain binds to the CIF150/hTAF II 150-binding domain, wherein if the first fusion protein comprises a DNA binding domain, then the second fusion protein comprises a transcriptional activating domain, wherein if the first fusion protein comprises a transcriptional activating domain, then the second fusion protein comprises a DNA binding domain, wherein the interaction of the first and second fusion proteins reconstitutes a sequence-specific transcription activating factor; and  
 iii) a reporter gene comprising a DNA sequence to which the DNA binding domain specifically binds; and  
   (b) measuring the expression of the reporter gene, wherein a test compound which decreases the expression of the reporter gene is a potential inducer of mitosis or cell cycle progression.