Table 3.
Analysis of nucleotide differences for individual MDIR domains
| A | |||||||||
| Type I and Type II | 1 | 2 | 3 | 4 | 5 | 6 | 7 | 8 | 9 |
| 1. MDIR1-IgD | |||||||||
| 2. MDIR2-IgD1 | 0.715 | ||||||||
| 3. MDIR3-IgD1 | 0.674 | 0.177 | |||||||
| 4. MDIR3-IgD2 | 0.683 | 0.172 | 0.027 | ||||||
| 5. MDIR5-IgD1 | 0.703 | 0.170 | 0.027 | 0.027 | |||||
| 6. MDIR4-IgD1 | 0.745 | 0.217 | 0.159 | 0.167 | 0.146 | ||||
| 7. MDIR4-IgD3 | 0.723 | 0.222 | 0.185 | 0.186 | 0.191 | 0.247 | |||
| 8. MDIR3-IgD4 | 0.685 | 0.191 | 0.141 | 0.139 | 0.171 | 0.198 | 0.115 | ||
| 9. MDIR4-IgD5 | 0.672 | 0.189 | 0.104 | 0.115 | 0.134 | 0.176 | 0.127 | 0.067 | |
| 10. MDIR5-IgD3 | 0.676 | 0.185 | 0.142 | 0.153 | 0.158 | 0.206 | 0.122 | 0.082 | 0.067 |
| Type III and Type IV | |||||||||
| 1 | 2 | 3 | 4 | 5 | 6 | 7 | |||
| 1. MDIR2-IgD2 | |||||||||
| 2. MDIR3-IgD5 | 0.245 | ||||||||
| 3. MDIR4-IgD6 | 0.264 | 0.131 | |||||||
| 4. MDIR5-IgD4 | 0.327 | 0.201 | 0.139 | ||||||
| 5. MDIR3-IgD3 | 0.318 | 0.399 | 0.405 | 0.418 | |||||
| 6. MDIR4-IgD2 | 0.327 | 0.417 | 0.420 | 0.421 | 0.051 | ||||
| 7. MDIR4-IgD4 | 0.275 | 0.389 | 0.385 | 0.368 | 0.078 | 0.077 | |||
| 8. MDIR5-IgD2 | 0.283 | 0.367 | 0.396 | 0.406 | 0.051 | 0.063 | 0.091 | ||
| B | |||||||||
| Type I and Type II | |||||||||
| 1 | 2 | 3 | 4 | 5 | 6 | 7 | 8 | 9 | |
| 1. MDIR1-IgD | |||||||||
| 2. MDIR2-IgD1 | 0.423 | ||||||||
| 3. MDIR3-IgD1 | 0.441 | 0.207 | |||||||
| 4. MDIR3-IgD2 | 0.443 | 0.187 | 0.060 | ||||||
| 5. MDIR5-IgD1 | 0.454 | 0.200 | 0.104 | 0.060 | |||||
| 6. MDIR4-IgD1 | 0.453 | 0.158 | 0.173 | 0.154 | 0.178 | ||||
| 7. MDIR4-IgD3 | 0.444 | 0.214 | 0.195 | 0.182 | 0.176 | 0.180 | |||
| 8. MDIR3-IgD4 | 0.443 | 0.185 | 0.171 | 0.154 | 0.152 | 0.161 | 0.101 | ||
| 9. MDIR4-IgD5 | 0.441 | 0.198 | 0.196 | 0.179 | 0.182 | 0.185 | 0.128 | 0.052 | |
| 10. MDIR5-IgD3 | 0.458 | 0.217 | 0.209 | 0.192 | 0.191 | 0.193 | 0.133 | 0.060 | 0.086 |
| Type III and Type IV | |||||||||
| 1 | 2 | 3 | 4 | 5 | 6 | 7 | |||
| 1. MDIR2-IgD2 | |||||||||
| 2. MDIR3-IgD5 | 0.218 | ||||||||
| 3. MDIR4-IgD6 | 0.186 | 0.071 | |||||||
| 4. MDIR5-IgD4 | 0.218 | 0.097 | 0.061 | ||||||
| 5. MDIR3-IgD3 | 0.290 | 0.298 | 0.266 | 0.279 | |||||
| 6. MDIR4-IgD2 | 0.260 | 0.278 | 0.247 | 0.273 | 0.071 | ||||
| 7. MDIR4-IgD4 | 0.256 | 0.293 | 0.268 | 0.292 | 0.133 | 0.089 | |||
| 8. MDIR5-IgD2 | 0.249 | 0.260 | 0.229 | 0.261 | 0.080 | 0.058 | 0.093 |
A The proportion of synonymous nucleotide differences per synonymous site (Ps) estimated by Nei and Gojobori’s method (Nei and Gojobori 1986). B The proportion of non-synonymous nucleotide differences per non-synonymous site (Pn)
From Table 3, the following points can be made: The theoretical maximum value of Ps is 0.75, if the frequencies of the four different types of nucleotide are assumed to be equal. Therefore, the MDIR1 and MDIR2-MDIR5 sequences appear to have been diverged for a long period of time, as the proportion of synonymous nucleotide differences per synonymous sites are close to saturation
MDIR3-IgD1 and MDIR3-IgD2 are results of recent duplications, as the proportion of synonymous nucleotide differences per synonymous site is small
Two distinct types of MDIR intermediate (I)-type IgSF domains emerge: (a) Type I: MDIR1-IgD; (b1) Type II subtype 1: MDIR2-IgD1, MDIR3-IgD1, MDIR3-IgD2, MDIR5-IgD1, MDIR4-IgD1; (b2) Type II, subtype 2: MDIR3-IgD4, MDIR4-IgD3, MDIR4-IgD5, MDIR5-IgD3
Two distinct C2-types emerge: Type III: MDIR2-IgD2, MDIR3-IgD5, MDIR4-IgD6, MDIR5-IgD4; Type IV: MDIR3-IgD3, MDIR4-IgD2, MDIR4-IgD4, MDIR5-IgD2
Within the MDIR domain type II, subtype 1 or type III categories, the respective domains of MDIR2 show higher differences from MDIR3-5, indicating the MDIR2 may have diverged first from MDIR3-5. The proportion of synonymous nucleotide differences per synonymous site for MDIR3-IgD1/IgD2 vs MDIR5-IgD1, MDIR3-IgD3 vs MDIR5-IgD2, MDIR3-IgD4 vs MDIR5-IgD3, MDIR3-IgD5 vs MDIR5-IgD4 are 0.027, 0.051, 0.082, and 0.201, respectively. Simple duplication of Ig domains of MDIRs does not explain the observation, implying instead an exon shuffling or gene conversion-like event between ancestors of MDIR3 and MDIR5