Novel mutations in the RING-finger domain of BRCA1 gene in clinically diagnosed breast cancer patients

  • 10 Accesses


In the present study, we investigated the frequency of BRCA1 gene mutations in 30 breast cancer (BC) patients of independent family history and 30 healthy control subjects. The immunohistochemistry (IHC) of BC patients showed duct cell carcinoma and distinct expression of the human epidermal growth factor receptor 2 (HER2). The genomic DNA was extracted from the BC patients and control subjects, the BRCA1 gene was PCR amplified and sequenced. The sequence analysis revealed that BRCA1 gene mutations were detected in 5/30 (16.6%) unrelated patients. One novel deleterious c.53delT mutation was detected in 3/30 (10%) unrelated patients leading to p.Met18Serfs*5 frame shift mutation in exon 2. Two patients 2/30 (6%) had novel c.297_301delinsCTCAA mutation in exon 5 leading to p.Leu99_Tyr101delinsPheSerAsn. Interestingly, the qRT-PCR analysis showed high expression of BRCA1 gene in all these patients having mutations compared with control subjects. Further, in silico analysis revealed loss of zinc-binding region of the RING-finger domain in BRCA1 structure due to these mutations, variable number of helices, helix–helix interactions, β-turns, and γ-turns were identified in the secondary structure, resulted in the formation of non-functional protein which is unable to activate BRCA1-associated genome surveillance complex (BASC) leading to uncontrolled cell proliferation. Moreover, the molecular dynamics (MD) simulations of mutated BRCA1 protein demonstrated extensive variations in the domain and non-domain regions compared with the wild-type structure as indicated by RMSD values. All these results conclusively explain that the c.53delT mutation may be the probable founder of deleterious mutation in this ethnic group.

This is a preview of subscription content, log in to check access.

Access options

Buy single article

Instant unlimited access to the full article PDF.

US$ 39.95

Price includes VAT for USA

Subscribe to journal

Immediate online access to all issues from 2019. Subscription will auto renew annually.

US$ 210

This is the net price. Taxes to be calculated in checkout.

Fig. 1
Fig. 2
Fig. 3
Fig. 4
Fig. 5
Fig. 6


  1. Bramucci E, Paiardini A, Bossa F, Pascarella S (2012) PyMod: sequence similarity searches, multiple sequence-structure alignments, and homology modeling within PyMOL. BMC Bioinform 13(Suppl4):S2

  2. Cortez D, Wang Y, Qin J, Elledge SJ (1999) Requirement of ATM-dependent phosphorylation of BRCA1 in the DNA damage response to double-strand breaks. Science 286:1162–1166

  3. Ellsworth RE, Decewicz DJ, Shriver CD, Ellsworth DL (2010) Breast cancer in the personal genomics era. Curr Genomics 11:146–161

  4. Ford D, Easton DF, Stratton M, Narod S, Goldgar D, Devilee P et al (1998) Genetic heterogeneity and penetrance analysis of the BRCA1 and BRCA2 genes in breast cancer families. The breast cancer linkage consortium. Am J Hum Genet 62:676–689

  5. Frank TS, Deffenbaugh AM, Reid JE, Hulick M, Ward BE, Lingenfelter B et al (2002) Clinical characteristics of individuals with germline mutations in BRCA1 and BRCA2: analysis of 10,000 individuals. J Clin Oncol 20:1480–1490

  6. Gatei M, Scott SP, Filippovitch I, Soronika N, Lavin MF et al (2000) Role for ATM in DNA damage-induced phosphorylation of BRCA1. Can Res 60:3299–3304

  7. Gudmundsdottir K, Ashworth A (2006) The roles of BRCA1 and BRCA2 and associated proteins in the maintenance of genomic stability. Oncogene 25:5864–5874

  8. Jamroz M, Kolinski A, Kmiecik S (2013) CABS-flex: Server for fast simulation of protein structure fluctuations. Nucleic Acids Res 41:W427–431

  9. Jensen RA, Thompson ME, Jetton TL, Szabo CI, van der Meer R, Helou B et al (1996) BRCA1 is secreted and exhibits properties of a granin. Nat Genet 12:303–308

  10. Kumar PS, Venkatesh K, Sowjenya G, Srikanth L, Sunitha MM, Prasad UV et al (2015) Mutations in exons 3 and 7 resulting in truncated expression of human ATP6V1B1 gene showing structural variations contributing to poor substrate binding-causative reason for distal renal tubular acidosis with sensorineural deafness. J Biomol Struct Dyn 33:2094–2103

  11. Laskowski RA (2001) PDBsum: summaries and analyses of PDB structures. Nucleic Acids Res 29:221–222

  12. Lee C, Banerjee T, Gillespie J, Ceravolo A, Parvinsmith MR, Starita LM et al (2015) Functional analysis of BARD1 missense variants in homology-directed repair of DNA double strand breaks. Hum Mutat 36:1205–1214

  13. Livak KJ, Schmittgen TD (2001) Analysis of relative gene expression data using real-time quantitative PCR and the 2(-delta delta C(T)) method. Methods 25:402–408

  14. Maresca L, Spugnesi L, Lodovichi S, Cozzani C, Naccarato AG, Tancredi M et al (2015) MSH2 role in BRCA1-driven tumorigenesis: a preliminary study in yeast and in human tumors from BRCA1-VUS carriers. Eur J Med Genet 58(10):531–539

  15. Miki Y, Swensen J, Shattuck-Eidens D, Futreal PA, Harshman K, Tavtigian S et al (1994) A strong candidate for the breast and ovarian cancer susceptibility gene BRCA1. Science 266:66–71

  16. Millot GA, Carvalho MA, Caputo SM, Vreeswijk MP, Brown MA, Webb M et al (2012) A guide for functional analysis of BRCA1 variants of uncertain significance. Hum Mutat 33:1526–1537

  17. Pasupuleti SK, Katari V, Lokanathan S, Uppu VP, Thummaginjala SS, Akkamgari RP et al (2014) Novel frame shift mutations ('A' deletion) observed in exon 9 of Wilms' tumor (WT1) gene in a patient reported with glomerulosclerosis. Gene 546:63–67

  18. Shin DS, Chahwan C, Huffman JL, Tainer JA (2004) Structure and function of the double-strand break repair machinery. DNA Repair (Amst) 3:863–873

  19. Thompson JD, Gibson TJ, Plewniak F, Jeanmougin F, Higgins DG (1997) The CLUSTAL_X windows interface: flexible strategies for multiple sequence alignment aided by quality analysis tools. Nucleic Acids Res 25:4876–4882

  20. Troudi W, Uhrhammer N, Sibille C, Dahan C, Mahfoudh W, Bouchlaka-Souissi C et al (2007) Contribution of the BRCA1 and BRCA2 mutations to breast cancer in Tunisia. J Hum Genet 52(11):915–920

  21. Vaidyanathan K, Kumar P, Reddy CO, Deshmane V, Somasundaramm K, Mukherjee G (2010) ErbB-2 expression and its association with other biological parameters of breast cancer among Indian women. Indian J Cancer 47:8–15

  22. van Gent DC, Hoeijmakers JH, Kanaar R (2001) Chromosomal stability and the DNA double-stranded break connection. Nat Rev Genet 2:196–206

  23. Venkatesh K, Srikanth L, Vengamma B, Chandrasekhar C, Prasad BC, Sarma PV (2015) In vitro transdifferentiation of human cultured CD34+ stem cells into oligodendrocyte precursors using thyroid hormones. Neurosci Lett 588:36–41

  24. Wooster R, Weber BL (2003) Breast and ovarian cancer. N Engl J Med 348:2339–2347

  25. Wu LC, Wang ZW, Tsan JT, Spillman MA, Phung A, Xu XL et al (1996) Identification of a RING protein that can interact in vivo with the BRCA1 gene product. Nat Genet 14:430–440

Download references


We sincerely acknowledge Sri Venkateswara Institute of Medical Sciences and University, Tirupati, Andhra Pradesh, India, for providing facilities to carry out this work.

Author information

Correspondence to Potukuchi Venkata Gurunadha Krishna Sarma.

Ethics declarations

Conflict of interest

All authors declare that there is no conflict of interest.

Rights and permissions

Reprints and Permissions

About this article

Verify currency and authenticity via CrossMark

Cite this article

Kumar, P.S., Srikanth, L., Reddy, K.S. et al. Novel mutations in the RING-finger domain of BRCA1 gene in clinically diagnosed breast cancer patients. 3 Biotech 10, 47 (2020) doi:10.1007/s13205-019-2037-5

Download citation


  • BRCA1 complex
  • RING-finger domain mutations
  • qRT-PCR
  • In silico analysis