3.2 Interaction of lamin A with telomeric complex
In order to confirm the bioinformatics data and to prove the hypothesis,
the genomic DNA was extracted from the human dermal fibroblast (HDF)
cells and restricted with specific restriction enzymes namely; Rsa I and
Hinf I for telomeric DNA coating experiments. Restricted genomic DNA has
intact telomeric repetitive sequences (TTAGGG)n. After coating of the
telomeric repetitive DNA, lamin A and trf2 specific antibodies were
added into protein lysate and determined the lamin A as well as trf2
binding capability. The data was shown in the (Figure 1C), which clearly
illustrates that lamin A binds strongly with the telomere repetitive DNA
sequences (TTAGGG)n. Similarly, as a positive control trf2 shows high
fluorescence intensity, which indicates that trf2 has strongly bound
with telomere repetitive DNA. Interestingly, the fluorescence intensity
of trf2 is higher than lamin A, which indicates that very short
percentage of lamin A amino acid residues may interact with the telomere
repetitive DNA sequences. Further to validate the data, we performed
co-immunoprecipitation.