CF Genetic Analysis Consortium NEWSLETTER #66-                                        

1. Summary of new CF mutations and DNA sequence polymorphisms:

Name of        Nucleotide      Exon    Consequence         Institute          Names of           
mutation       change                                                         Contributors       
I1269N         T->A at 3938    20      Ile->Asn at 1269    Institute of       McDowell T,        
                                                           Molecular          Shackleton S,      
                                                           Medicine, Oxford   Harris A (Mar 28)  
4005+28insA    insertion of    intron  polymorphism?       same as above      McDowell et al.    
               A after         20                                             (Mar 28)           
Note:  The above sequence alterations were found by SSCP analysis.  I1269N was found in 2                    
sisters with [[Delta]]F508 on their aother allele.  The mutation destroys a TaqI                             
restriction site. Contact: Dr. Ann Harris; Address: Pediatric Molecular Genetics, Institute                  
of Molecular Medicine, John Radcliffe Hospital, Headington, Oxford OX3 9DU, England;                         
Telephone: +44 (0865) 222341/222340; FAX: +44 (0865) 220479; E-mail:                                                                                                                     
S466L          C->T at 1529    9       Ser->Leu at 466     Institut           Costes B, Ghanem   
                                       (CBAVD)             National de la     N, Girodon E,      
                                                           Sante et de la     Goossens M (Apr    
                                                           Recherche          4)                 
Note:  The above mutation was detected by DGGE and identified by direct sequencing in an                     
infertile man with isolated CBAVD. Contact: Prof. Michel Goossens; Address: INSERM U.91,                     
CHU Henri Mondor, 51 Av. du Maréchal de Lattre de Tassigny, 94010 CRETEIL, France;                           
Telephone: +33 (1) 49 81 28 60; FAX: +33 (1) 49 81 28 42                                                     
N1303I         A->T at 4040    21      Asn->Ile at 1303    Free University    Lissens W,         
                                                           Hospital Brussels  Bonuelle M,        
                                                                              Seneca S,          
                                                                              Liebaers (Apr      
                                                                              21); also Férec    
                                                                              et al. (Jun 14)    
Note:  N1303I was found in a normal individual with no family history of CF.  It was                         
detected by using a modified primer designed to detect the N1303K mutation after PCR in                      
combination with primer 21i5 and restriction digestion with BsrI.  This individual was                       
negative for N1303K by reverse dot blot analysis but heterozygous after digestion with                       
BsrI.  The change was then detected by direct sequencing of exon 21 after amplification                      
with 21i5 and 21i3. Contact: Prof. Inge Liebaers; Address: Center for Medical Genetics,                      
Free University Hospital Brussels, Laarbeeklaan 101, 1090 Brussels, Belgium;  Telephone:                     
+32 (02) 477.60.71;  FAX: +32 (02) 477.58.00                                                                 
R764X          C->T at 2422    13      Arg->Stop at 764    Johns Hopkins      Macek MJr, Egan    
                                                           Medical            ME, Mackova A,     
                                                           Institutions       Cutting GR (Apr    
Note:  The above was deleted in a 32-year old African-American female CF patient whose                       
other mutation was [[Delta]]F508.  She was diagnosed at age 11 with repeated respiratory                     
symptoms but remained pancreatic sufficient thus far. Contact: Dr. Garry R. Cutting;                         
Address: Center for Medical Genetics, 600 N. Wolfe Street, CMSC 1004, Baltimore, MD                          
21287-3914; Telephone: +1 (410) 614-0211; FAX: +1 (410) 955-0484; e-mail:                                                                                                          
R792G          C->G at 2506    13      Arg->Gly at 792     National           Glavac D,          
                                                           Institute of       Ravnik-Glavac M,   
                                                           Chemistry,         Dean M (Apr 25)    
R766M          G->T at 2429    13      Arg->Met at 766     same as above      Glavac et          
                                                                              al.(May 3)         
Note:  R792P was detected by SSCP and heteroduplex analysis followed by direct sequencing.                   
R766M was identified by non-radioactive SSCP and direct sequencing; it creates a FokI                        
restriction site. Contact:  Damjan Glavac; Address: National Institute of Chemistry,                         
SI-61115 Ljubljana, Hajdrihova 19, Slovenia, P.O.B. 30; Telephone: (+386 61) 123-20-61;                      
FAX: (+386 61) 125-92-44/125-70-69                                                                           
622-1G->A      G->A at 622-1   intron  splice mutation     The Hospital for   Zielenski J,       
                               4                           Sick Children,     Markiewicz D,      
                                                           Toronto            Tsui L-C, Casals   
                                                                              T, Ramos MD,       
                                                                              Estivill X, Bal    
                                                                              J, Mazurczak T     
                                                                              (Apr 28)           
Note:  The above mutation was detected was detected by heteroduplex and DGGE analyses.  It                   
was found in 2 Polish CF patients.  The first patient was diagnosed with PI, moderate lung                   
disease, sweat chloride of 90.7-141.2 mEq/l, and with second mutation being 2143delT.  The                   
second patient was also with PI, moderate to severe lung disease, sweat chloride of 76-108,                  
and with unknown second mutation. Contact: Dr. Lap-Chee Tsui or Julian Zielenski;  Address:                  
Department of Genetics, The Hospital for Sick Children, 555 University Avenue, Toronto,                      
Ontario, M5G 1X8 CANADA; Telephone: (416) 813 6365   FAX: (416) 813 4931; E-Mail:                                                                                                           
I539T          T->C at 1748    11      Ile->Thr at 539     Laboratoire de     Chomel J-C,        
                                                           Génétique          Kitzis A (May 19)  
                                                           Cellulaire et                         
                                                           Poitiers, France                      
Note:  The above mutation was detected by DGGE with chemical clamps and characterized by                     
direct sequencing. Contact: Jean-Claude Chomel; Address: Centre Hospitalier Universitaire                    
de Poitiers, Hôpital Jean-Benard- La Milétrie- BP 577-86021, Pointiers Cedex; Telephone:                     
+33 /57.94 /57.95; FAX: +33                                                          
Y247X          C->G at 873     6a      Tyr->Stop at 247    Athens             Tzetis M,          
                                                           University First   Antoniadi T,       
                                                           Dept. of           Kanavakis E (May   
                                                           Pediatrics         30)                
2752-26A->G    A->G at         intron  splice mutation     same as above      Tzetis et al       
               2752-26         14a                                            (May 30)           
Note:  Y247X was detected by DGGE and confirmed by direct sequencing.  It was found in a CF                  
adult, currently 21 years old, of Greek origin out of 472 CF alleles tested.  The patient's                  
other CF allele is [[Delta]]F508.  The patient has pancreatic insufficient since birth                       
sweat chloride of 81.2 mEq/L, chronic cough and obstructive lung disease. 2752-26A->G was                    
detected by DGGE and identified by direct sequencing out of 472 CF alleles tested.  The                      
patient, of Greek origin, is currently 2.5 years of age.  The patient's DNA was also                         
analyzed by DGGE for exons 3, 4, 5, 6a, 7, 8, 10, 11, 12, 14a, 14b, 15, 17b, 18, 19, 20, 21                  
and for mutation 3849+10kbC->T with HphI, but no other alteration could be detected.                         
2752-26A->G probably creates an alternative splice site that competes with the normal                        
acceptor site, causing reduction of full length mRNA synthesis. Contact: Dr. med. Emmanouil                  
Kanavakis;  Address: Anthens University, First Dept. of Pediatrics, Unit of Molecular                        
Medicine, St. Sophia Children's Hospital, Athens 11527, Greece; Telephone/FAX: (031)                         
7795762; E-Mail:                                                             
3600+5G->A     G->A at 3600+5  intron  splice mutation?    Groupe             Bienvenu T,        
                               18                          Hospitalier        Bousquet S,        
                                                           Cochin, Paris      Beldjord C,        
                                                                              Kaplan JC (Jun 8)  
3384A/G        A or G at 3384  17b     No change (Leu at   Groupe             Bienvenu et al.    
                                       1084)               Hospitalier        (Jun 16)           
                                                           Cochin, Paris                         
Note:  The above 2 mutations were detected by DGGE using chemical clamps and identified by                   
direct sequencing.  3600+5G->A has been found only once among 50 non-[[Delta]]F508 CF                        
chromosome and 50 non-CF chromosomes, whereas 3394A/G, among 100 non-[[Delta]]F508 CF                        
chromosome and 100 non-CF chromosomes. Contact: Jean Claude Kaplan; Address: Groupe                          
Hospitalier Cochin, 27 rue de fg Saint-Jacques, 75679 Paris Cedex 14; Telephone: +33 (1)            ; FAX: +33 (1)                                                                        
237insA        insertion of    2       frameshift          Centre de          Férec C,           
               A after 237                                 Transfusion        Verlingue C,       
                                                           Sanguine et de     Quere I,           
                                                           Biogénétique,      Raguenes O,        
                                                           Brest, France      Audrezet M-P,      
                                                                              Mercier B (Jun 8)  
G550R          G->A at 1780    11      Gly->Arg at 550     same as above      Férec et al.       
                                                                              (Jun 8)            
N1303I         A->T at 4040    21      Asn->Ile at 1303    same as above      Férec et al.       
                                                                              (Jun 14)           
Note:  The above mutations were detected by DGGE and identified direct sequencing. Contact:                  
Dr. Claude Férec; Address: Centre de Biogénétique, Centre de Transfusion Sanguine et de                      
Biogénétique, 46 Rue Félix Le Dantec - B.P. 454 - 29275 BREST Cédex, France; Telephone: +33         ; FAX: +33                                                                            
[[Delta]]D192  deletion of     5       deletion of Asp     Hôpital            Feldmann D,        
               TGA or GAT              at 192              D'enfants          Magnier C,         
               from 706 or                                 Armand-Trousseau   Chauve C, Laroze   
               707                                                            F, Aymard P,       
                                                                              Grimfeld A (Jun    
Note:  The mutation was detected by DGGE and identified by direct sequencing in 2 siblings                   
with CF.  Their other CF allele is 711+1G->T.  They are both pancreatic insufficient with                    
sweat chloride of 121 and 95 mEq/L, respectively. Contact: Dr. Delphine Feldmann; Address:                   
Laboratoire de Biochimie, Hôpital A. Trousseau, 26 avenue du Dr A. Netter, 75571 Paris                       
Cédex 12, France;  Telephone: +33; FAX: +33                                          
2839T/C        T or C at 2839  15      Tyr or His at 903   Laikon General     Balassopoulou A,   
                                                           Hospital, Athens   Hatzipanaiotou N   
                                                                              (Jun 30)           
Note:  The above polymorphism was detected by DGGE and direct sequencing.  It was found on                   
the normal chromosome of the father of a CF patient, and was not detected in 70 normal and                   
150 CF chromosomes. Contact: Dr. Angeliki Balassopoulou; Address: Center if Thalassemias,                    
Unit of Prenatal Diagnosis, 16 Sevastoupoleos Street, Ampelokipi 11526, Athens, Greece;                      
Telephone:  +30 1-77.89.476 ; FAX: +30 1-77.57.442                                                           
347delC        deletion of C   3       frameshift          Hôpital Debrousse  Chevalier-Porst    
               at 347                                                         F, Bozon D (Jun    
del40kb        deletion of     4-10    large deletion      same as above      Chevalier-Porst    
               exons 4-10                                                     & Bozon (Jun 30)   
Comment:  347delC was found in one French CF patient, with the other mutation being N1303K.                  
del40kb was detected by PFGE in a French patient with [[Delta]]F508 on the other                             
chromosome. Contact: Dominique Bozon; Address: Biochimie Bât D, Hôpital Debrousse, 29 rue                    
Soeur Bouvier, 69322 Lyon, Cedex 05, France; Telephone: +33; FAX: +33 72.                        
G480D          G->A at 1570    10      Gly->Asp at 480     Royal Manchester   Hawworth A,        
                                                           Children's         Malone G,          
                                                           Hospital, England  Schwarz M (Jul 3)  
1058delC       deletion of C   7       frameshift          same as above      Malone G,          
               at 1058                                                        Haworth A,         
                                                                              Schwarz M (Jul 3)  
621+2T->C      T->C at 621+2   intron  splice mutation     same as above      Schwarz M,         
                               4                                              Malone G,          
                                                                              Hawaorth A (Jul    
H620P          A->C at 1991    13      His->Pro at 620     same as above      Haworth et al.     
                                                                              (Jul 10)           
Comment:  The above 4 mutations were detected by SSCP and identified by direct DNA                           
sequencing.  G480D was found in a CF patient from the West Midlands, who had meconium ileus                  
and whose other chromosome carries [[Delta]]F508; it was seen only once in 100                               
non-[[Delta]]F508 chromosome screened.  1058delC was detected only once in 200                               
non-[[Delta]]F508 chromosomes screened; the patient has [[Delta]]F508 on the other                           
chromosome.  621+2T->C was found in a Pakistani CF patient from the North-West of England;                   
the patient is homozygous for this mutation from a consanguineous partnership; the mutation                  
was found only once (although in homozygous form) in 20 Pakistani CF chromosomes.  H620P                     
was found in a 15-year old male CF patient who has mild CF and who is pancreatic                             
sufficient.  His parents were unavailable for testing but one is Caucasian and the other is                  
Asian.  His mother's mutation is R1158X, which this investigators have seen in one Arabic                    
and one Greek patient.  H620P was seen only once in 100 non-[[Delta]]F508 chromosomes                        
screened. Contact: Dr. Martin Schwarz; Address: Regional Molecular Genetics Laboratory,                      
Royal Manchester Children's Hospital, Hospital Road, Pendlebury, Manchester M27 4HA;                         
Telephone: 0161-794 4696; FAX: 0617272328                                                                    

2. Macek MJr, Egan ME, Mackova A, and Cutting GR (Apr 24) noted that the correct name for K166Q (NL#62) should be K166E; resequencing of 1342-1G->C (intron 8) showed that it should be 1342-2delAG.

3. I have received several positive (and no negative) comments regarding the new Newsletter format and the INTERNET display. We have also noted frequent login from many viewers, some non-consoritum members, indicating that the database is quite popular. Again, the World Wide Web address is <>. The number is still a temporary one for this experiment. Link-up with Genome Database and other network is being explored. We are also working on automatic e-mail retrieval, which is probably more accessible for most members.

Please drop me note by e-mail <> that you have seen it and give me any suggestions regarding future improvements.