Analysis of sperm nuclear protein gene polymorphisms and DNA integrity in infertile men

Figures & data

Figure 1. Schematic representation of PRM1 and PRM2 gene nucleotide sequence. The blue colored regions are introns. Nucleotide changes are green and the variant is shown just above in brown with their respective position. The corresponding amino acid change is shown below the wild type in red.

Figure 2. Sequence electrophoreogram of the SNP variant. The nucleotide variant SNP c443C > A in PRM2 gene is resolved. A) represents wild type sequence (C/C); B) represents sequence showing homozygous mutant (A/A).

Table 1. Sperm PRM and TNP genotypic and allelic frequency in oligozoospermic infertile and control men.

Gene	Nt position/ Nt change	Region	NCB ID	Codon change	Amino acid change	Infertile patient (n = 100)		P^a	Controls (n = 100)		P^b
						Allele frequency (%)	Genotype frequency (%)		Allele frequency (%)	Genotype frequency (%)
PRM1	230 C → A	Exon 2	rs737008	CGA → AGA	R47R	C: 0.66A:0.34	CC-0.56CA-0.20AA-0.24	NS	C:0.60A:0.40	CC-0.48CA-0.24AA-0.28	NS
PRM2	298 G → C	Intron 1	rs1646022	None	None	G:1.0C: 0.0	GG- 1.0GC-0.0CC-0.0	NS	G:0.98C:0.02	GG-0.98GC- 0.00CC-0.02	NS
PRM2	373 C → A	Intron 1	rs2070923	None	None	C:0.65A:0.35	CC- 0.55CA-0.20AA- 0.25	NS	C:0.60A:0.40	CC- 0.60CA-0.00AA-0.40	NS
PRM2	443 C → A	Exon 2	Novel^c	ACC → AAC	T94N	C:0.99A:0.01	CC-0.99CA-0.00AA-0.01	NS	C:1.0A:0.0	CC-1.0CA- 0.0AA- 0.0	NS
TNP1	214 T → C	Intron	rs62180545	None	None	T:0.93C:0.07	TT-0.85TC-0.15CC-0.00	NS	T:1.0C:0.0	TT-1.0TC-0.0CC-0.0	< 0.05
TNP1	Ins T 396-397 3′UTR	UTR	Novel^d	-	-	-	-	-	-	-	-
TNP2	391 C → T	Exon-1	rs11640138	CGC → TGC	R101C	C:0.40T:0.60	CC-0.40CT-0.00TT- 0.60	NS	C:0.43T:0.57	CC-0.43CT-0.00TT-0.57	NS

P^a: significance of allelic frequency between patients and controls

P^b: significance of genoptype frequency between patients and controls

^c: Gene bank ID- GU190363; ^d:- Gene bank ID- GU190364

Figure 3. Schematic representations of TNP1 and TNP2 genes showing the location of the SNPs. Blue-colored regions are introns. Nucleotide changes are green and the variant is shown just above in brown with their respective position. The corresponding amino acid change is shown below the wild type in red. The non coding exon region is shown underlined.

Figure 4. Sequence electrophoreogram of the SNP variant. The insertion c. Ins T 396-397 3′UTR TNP1 gene is highlighted. A) represents sequence showing homozygous wild-type (GGTG); B) represents sequence showing insertion (GGTTG).

Figure 5. SCSA pseudocolor dot plot of semen sample presenting with c443C > A mutation. FL 3-H -axis represents fragmented DNA and FL 1-H- axis represents native DNA. HDS: high DNA stainability cells in the semen.

Figure 6. SCSA pseudocolor dot plot of semen sample presenting with c.396_397InsT mutation. FL 3-H -axis represents fragmented DNA and FL 1-H- axis represents native DNA. HDS: high DNA stainability cells in the semen.