Heregulin-like Factor

Abstract

The present invention relates to a novel HLF protein which is a member of the heregulin family. In particular, isolated nucleic acid molecules are provided encoding the human HLF protein. HLF polypeptides are also provided as are vectors, host cells and recombinant methods for producing the same. The invention further relates to screening methods for identifying agonists and antagonists of HLF activity. Also provided are diagnostic methods for detecting disorders of the regulation of cell growth and therapeutic methods for treating disorders of the regulation of cell growth.

1 . An isolated nucleic acid molecule nucleic acid molecule comprising a polynucleotide having a nucleotide sequence at least 95% identical to a sequence selected from the group consisting of:
 (a) a nucleotide sequence encoding the HLF polypeptide having the amino acid sequence at positions 1 to 157 of SEQ ID NO:2) or the complete amino acid sequence encoded by the cDNA clone contained in ATCC Deposit No. 209123;   (b) a nucleotide sequence encoding the predicted extracellular domain of the HLF polypeptide having the amino acid sequence in SEQ ID NO:2 (i.e., positions 1 to 101 of SEQ ID NO:2) or as encoded by the cDNA clone contained in ATCC Deposit No. 209123;   (c) a nucleotide sequence encoding the predicted transmembrane domain of the HLF polypeptide having the amino acid sequence in SEQ ID NO:2 (i.e., positions 102 to 121 of SEQ ID NO:2) or as encoded by the cDNA clone contained in ATCC Deposit No. 209123;   (d) a nucleotide sequence encoding the predicted intracellular domain of the HLF polypeptide having the amino acid sequence in SEQ ID NO:2 (i.e., positions 122 to 157 of SEQ ID NO:2) or as encoded by the cDNA clone contained in ATCC Deposit No. 209123;   (e) a nucleotide sequence encoding a soluble HLF polypeptide having the extracellular and intracellular domains but lacking the transmembrane domain;   (f) the nucleotide sequence of clone HAGFE38R (SEQ ID NO:4);   (g) the nucleotide sequence of a portion of the sequence shown in  FIGS. 1A and 1B  (SEQ ID NO: 1) wherein said portion comprises at least 50 contiguous nucleotides from nucleotide about 1 to about 220 and from about 400 to 2199;   (h) the nucleotide sequence of a portion of the sequence shown in  FIGS. 1A and 1B  (SEQ ID NO:1) wherein said portion consists of residues 1 to 2199, 1 to 1500, 1 to 1000, 1 to 500, 1 to 250, 250 to 2199, 250 to 1500, 250 to 1000, 250 to 500, 500 to 2199, 500 to 1500, 500 to 1000, 1000 to 2199, and 1000 to 1500; and   (i) a nucleotide sequence complementary to any of the nucleotide sequences in (a) through (e) above.   
     
     
         2 . The nucleic acid molecule of  claim 1  wherein said polynucleotide has the complete nucleotide sequence in  FIGS. 1A and 1B  (SEQ ID NO: 1). 
     
     
         3 . The nucleic acid molecule of  claim 1  wherein said polynucleotide has the nucleotide sequence in  FIGS. 1A and 1B  (SEQ ID NO: 1) encoding the HLF polypeptide having the amino acid sequence in positions 1 to 157 of SEQ ID NO:2. 
     
     
         4 . The nucleic acid molecule of  claim 1  wherein said polynucleotide has the nucleotide sequence in  FIGS. 1A and 1B  (SEQ ID NO: 1) encoding the extracellular domain of the HLF polypeptide having the amino acid sequence from about 1 to about 101 in SEQ ID NO:2. 
     
     
         5 . The nucleic acid molecule of  claim 1  wherein said polynucleotide has the complete nucleotide sequence of the cDNA clone contained in ATCC Deposit No. 209123. 
     
     
         6 . The nucleic acid molecule of  claim 1  wherein said polynucleotide has the nucleotide sequence encoding the HLF polypeptide having the complete amino acid sequence encoded by the cDNA clone contained in ATCC Deposit No. 209123. 
     
     
         7 . The nucleic acid molecule of  claim 1  wherein said polynucleotide has the nucleotide sequence encoding the extracellular domain of the HLF polypeptide having the amino acid sequence encoded by the cDNA clone contained in ATCC Deposit No. 209123. 
     
     
         8 . An isolated nucleic acid molecule comprising a polynucleotide which hybridizes under stringent hybridization conditions to a polynucleotide having a nucleotide sequence identical to a nucleotide sequence in (a), (b), (c), (d), or (e) of  claim 1  wherein said polynucleotide which hybridizes does not hybridize under stringent hybridization conditions to a polynucleotide having a nucleotide sequence consisting of only A residues or of only T residues. 
     
     
         9 . An isolated nucleic acid molecule comprising a polynucleotide which encodes the amino acid sequence of an epitope-bearing portion of a HLF polypeptide having an amino acid sequence in (a), (b), (c), (d), or (e) of  claim 1 . 
     
     
         10 . The isolated nucleic acid molecule of  claim 9 , which encodes an epitope-bearing portion of a HLF polypeptide wherein the amino acid sequence of said portion is selected from the group of sequences in SEQ ID NO:2 consisting of: about Ser-1 to about Thr-8, about Thr-9 to about Lys-18, about Thr-23 to about His-31, about Phe-32 to about Leu-40, about Cys-43 to about Val-51, about Thr-56 to about Tyr-68, about Gln-75 to about Leu-84, about Tyr-126 to about Ala-135, about Ser-137 to about Leu-146, and about Ser-148 to about Lys-157. 
     
     
         11 . A method for making a recombinant vector comprising inserting an isolated nucleic acid molecule of  claim 1  into a vector. 
     
     
         12 . A recombinant vector produced by the method of  claim 11 . 
     
     
         13 . A method of making a recombinant host cell comprising introducing the recombinant vector of  claim 12  into a host cell. 
     
     
         14 . A recombinant host cell produced by the method of  claim 13 . 
     
     
         15 . A recombinant method for producing a HLF polypeptide, comprising culturing the recombinant host cell of  claim 14  under conditions such that said polypeptide is expressed and recovering said polypeptide. 
     
     
         16 . An isolated HLF polypeptide comprising an amino acid sequence at least 95% identical to a sequence selected from the group consisting of:
 (a) the amino acid sequence of the HLF polypeptide having the complete amino acid sequence shown in SEQ ID NO:2 (i.e., positions 1 to 157 of SEQ ID NO:2) or the complete amino acid sequence encoded by the cDNA clone contained in the ATCC Deposit No. 209123;   (b) the amino acid sequence of the predicted extracellular domain of the HLF polypeptide having the amino acid sequence shown in SEQ ID NO:2 (i.e., positions 1 to 101 of SEQ ID NO:2) or as encoded by the cDNA clone contained in the ATCC Deposit No. 209123;   (c) the amino acid sequence of the predicted transmembrane domain of the HLF polypeptide having the amino acid sequence shown in SEQ ID NO:2 (i.e., positions 102 to 121 of SEQ ID NO:2) or as encoded by the cDNA clone contained in the ATCC Deposit No. 209123;   (d) the amino acid sequence of the predicted intracellular domain of the HLF polypeptide having the amino acid sequence shown in SEQ ID NO:2 (i.e., positions 122 to 157 of SEQ ID NO:2) or as encoded by the cDNA clone contained in the ATCC Deposit No. 209123; and   (e) the amino acid sequence of a soluble HLF polypeptide having the extracellular and intracellular domains but lacking the transmembrane domain.   
     
     
         17 . The isolated HLF polypeptide of  claim 16 , which comprises an epitope-bearing portion of the HLF protein, wherein said portion is selected from the group consisting of: a polypeptide comprising amino acid residues from about Ser-1 to about Thr-8, about Thr-9 to about Lys-18, about Thr-23 to about His-31, about Phe-32 to about Leu-40, about Cys-43 to about Val-51, about Thr-56 to about Tyr-68, about Gln-75 to about Leu-84, about Tyr-126 to about Ala-135, about Ser-137 to about Leu-146, and about Ser-148 to about Lys-157 of SEQ ID NO:2. 
     
     
         18 . An isolated antibody that binds specifically to a HLF polypeptide of  claim 16 . 
     
     
         19 . A method for diagnosing cancer in a human comprising,
 (a) assaying HLF gene expression level in cells or body fluid of an individual; and   (b) comparing the HLF gene expression level with a standard HLF gene expression level, whereby an increase or decrease in the assayed HLF gene expression level compared to the standard expression level is indicative of cancer in the tissue type assayed.