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) 4005+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: aharris@molbiol.ox.ac.uk 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) Medicale 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 24) 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: mmacek@welchlink.welch.jhu.edu R792G C->G at 2506 13 Arg->Gly at 792 National Glavac D, Institute of Ravnik-Glavac M, Chemistry, Dean M (Apr 25) Slovenia 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: cfdata@sickkids.on.ca I539T T->C at 1748 11 Ile->Thr at 539 Laboratoire de Chomel J-C, Génétique Kitzis A (May 19) Cellulaire et Moléculaire 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 49.44.39.03 /57.94 /57.95; FAX: +33 49.44.39.12 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: ekanavak@atlas.uoa.ariadne-t.gr 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) 42.34.12.12; FAX: +33 (1) 44.41.15.22 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 98.44.50.64; FAX: +33 98.43.05.55 [[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 13) 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 44.73.68.67; FAX: +33 44.73.66.87 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 30) 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 72.38.57.21; FAX: +33 72. 38.58.84 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 6) 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 <http://199.0.26.114/>. 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 <cfdata@sickkids.on.ca> that you have seen it and give me any suggestions regarding future improvements.
Regards,